SNAPs与吗啡依赖关系的初步实验研究
|
摘要
阿片依赖是一种慢性复发性脑病,以强迫用药、不断增加药物摄入量和停药时出现戒断综合征为主要特征。目前阿片依赖发生的确切机制还不清楚。大量研究表明,突触可塑性的改变在阿片依赖过程中起了重要作用。这些变化包括突触前神经递质的释放和突触后受体的信号转导。以往研究发现,吗啡可以通过调节突触前影响神经递质释放的分子影响递质释放。
在神经递质的释放过程中,囊泡膜和质膜的融合是影响递质释放的关键步骤。可溶性NSF附着蛋白(Soluble NSF attachment proteins,SNAPs)是一类在膜融合过程中发挥重要作用的蛋白质分子,介导了囊泡膜和质膜的融合,在哺乳动物中存在α-,β-,和γ- 3种亚型。鉴于SNAPs在膜融合过程中发挥的重要作用,SNAPs参与许多重要的生理进程,包括调节钙依赖的胞吐机制、刺激胰岛β细胞分泌胰岛素、增加肺泡II型细胞表面活性物质的分泌、参与有丝分裂过程中核膜的形成等。
虽然大量研究表明SNAPs在神经递质释放的膜融合过程中起重要作用,但是SNAPs是否参与了阿片依赖过程中突触前神经可塑性的发生目前尚未见报道。为了揭示SNAPs与阿片依赖的关系,我们首先在大鼠吗啡依赖及戒断模型上观察了与神经可塑性关系密切的脑区伏隔核(nucleus accumbens,NAc)、尾壳核(caudate putamen,CPu)及海马(hippocampus,Hip)中SNAPs表达的变化,初步探讨SNAPs和吗啡依赖的关系。研究采用成年雄性Wistar大鼠,随机分为对照组、吗啡组和戒断不同时间组。吗啡组大鼠背部皮下注射吗啡8d,每日3次,剂量递增,对照组大鼠注射等体积生理盐水。吗啡组末次注射吗啡4 h后处死,断头取脑。自然戒断组分别在戒断不同时间处死动物。各组均设平行对照。利用RT-PCR和Western blot技术分别检测各脑区SNAPs的mRNA和蛋白表达水平。结果发现,与对照组相比,慢性吗啡依赖大鼠CPu内γ-SNAP的mRNA和蛋白表达水平均上调25%左右(P<0.01) (图1.4,图1.9),NAc及Hip脑区则无明显变化,自然戒断d2、d3、d7组亦未观察到γ-SNAP明显的表达改变(图1.8,图1.10,表1.1,表1.3)。α-SNAP和β-SNAP在吗啡依赖和自然戒断状态下3个被检脑区(NAc、CPu、Hip)均未检测到明显的表达变化(图1.2,图1.3,图1.5,图1.6,图1.7,表1.1,表1.2)。以上结果说明:慢性吗啡依赖可增加CPu内γ-SNAP的表达,对α-SNAP和β-SNAP的表达没有影响,提示SNAPs 3种亚型在慢性吗啡依赖过程中行使不同的功能,可能与特定神经递质的分泌有关。慢性吗啡依赖及戒断引起的突触前神经递质释放改变可能不是由α-SNAP和β-SNAP表达量变化介导的,其具体机制可能与其内在活性变化或蛋白在细胞内的分布有关。
为了进一步研究SNAPs与阿片依赖的关系,我们选择了一种与神经递质释放关系最为明确的亚型—α-SNAP进行研究。已有大量研究报道α-SNAP参与钙依赖的胞吐作用,该作用在神经递质释放过程中的囊泡膜与质膜的融合步骤中是至关重要的。且相关研究报道α-SNAP通过刺激NSF的ATP酶活性来发挥其在递质释放中的作用,α-SNAP对NSF的ATP酶刺激作用明显强于γ-SNAP。因此,虽然我们在动物模型上未检测到α-SNAP的蛋白表达变化,但由于其在递质释放中的重要作用,我们依然选择这种亚型作为研究对象。为了进一步研究α-SNAP的功能与吗啡依赖的关系,选择一个良好的、调控模式相对简单和直接的细胞模型是十分必要的。该细胞模型必须能够满足以下条件:(1)表达阿片受体;(2)拥有类似成熟神经内分泌细胞的一些特性,能释放与阿片依赖有关的神经递质,如单胺类、谷氨酸(glutamate,Glu)、γ-氨基丁酸(γ-aminobutyric acid ,GABA)等。经过初步筛选,我们选择了分化的人神经母细胞瘤细胞株SH-SY5Y作为研究对象。据研究报道,SH-SY5Y细胞株经过维甲酸(retinoic acid,RA)分化后可表现出许多类似成熟神经元的特性,如神经突起变长、电刺激的兴奋性增加、神经元特异性烯醇化酶(neuron specific enolase,NSE)表达增加、神经分泌颗粒增多等。
在研究中我们发现,RA分化6天的SH-SY5Y细胞可形成较长的神经突起(图2.1),免疫印迹分析检测到分化后的细胞内NSE表达增加(图2.2)。另外,在我们的研究中亦发现了RA分化后的细胞μ阿片受体(μopioid receptor,MOR)表达上调,与以往研究相一致(图2.3)。这些特性都为我们研究阿片依赖的分子机制提供了有力的条件,尤其是与突触前神经递质释放有关的分子机制。为了研究与阿片依赖有关的分子机制,我们首先需要在SH-SY5Y细胞上建立一个阿片依赖的细胞模型。阿片长期暴露后给予纳络酮催促戒断出现cAMP超射常常被作为评定阿片依赖模型建立是否成功的一个重要指标。本研究采用免疫竞争结合法,测定了吗啡处理前后细胞内cAMP含量的变化,发现100μM吗啡作用分化SH-SY5Y细胞24h后给予纳络酮催促戒断可形成明显的cAMP超射,cAMP含量约为催促前的2.36倍(图2.5),证实了分化SH-SY5Y细胞上吗啡依赖的细胞模型建立是成功的。
为了进一步确定吗啡处理过程中神经递质的释放情况,我们采用高效液相-电化学检测法(high performance liquid chromatography- electrochemical detection,HPLC-ECD)测定了急性吗啡作用及慢性吗啡作用纳络酮催促戒断后不同时间SH-SY5Y细胞培养上清液中单胺类神经递质的含量,探讨吗啡处理与神经递质释放的关系。结果显示,在急性吗啡作用下,单胺类神经递质,包括去甲肾上腺素(norepinephrine,NE)、多巴胺(dopamine,DA)、5-羟色胺(5-hydroxytryptamine,5-HT)的释放均受到普遍抑制(表3.1)。100μM吗啡孵育SH-SY5Y细胞10min~1h,细胞培养上清内NE,DA和5-HT及其代谢产物3,4-二羟基苯乙酸(3,4-dihydroxy- phenylacetic acid,DOPAC) (DA代谢产物)和5-羟基吲哚乙酸(5-hydroxyindolacetic acid,5-HIAA )(5-HT代谢产物)均显著下降,其中NE含量从16.97 ng/g蛋白下降至5.34 ng/g蛋白,下降幅度以吗啡作用10min内下降最为明显(P <0.01),在吗啡继续作用的50min内一直保持在较低水平(图3.4)。DA的变化趋势与NE类似,在吗啡作用的最初10min亦出现明显下降,而且下降幅度在2/3以上(P <0.01) (图3.6)。虽然5-HT在吗啡急性作用的早期出现中等程度的下降,但在吗啡继续作用的50min内,5-HT含量继续降低,至吗啡作用后1h达到基础水平的1/3左右(P<0.01)(图3.8)。与吗啡处理前相比,代谢产物DOPAC和5-HIAA均出现不同程度的降低(P<0.01) (图3.5,图3.7)。这些结果表明,急性吗啡作用可以显著抑制SH-SY5Y细胞单胺类神经递质的释放。
在给予100μM吗啡孵育细胞24h后用100μM纳络酮催促戒断测定SH-SY5Y细胞上清单胺类神经递质的含量,结果发现:NE、DA和5-HT及其代谢产物DOPAC和5-HIAA均有不同程度的增加(表3.2)。NE的含量在戒断20 min后由戒断前7.59 ng/g蛋白增加至12.74 ng/g蛋白,增加50%左右,与催促前相比有显著性差异(P <0.01),在戒断40min时恢复至戒断前水平(图3.9)。与NE相比,DA的变化趋势更加明显,DA在戒断的10min即有增加趋势,但与催促前相比无显著性差异(P> 0.05),在戒断20min时达最高值,约为催促前基础水平的2倍(P <0.05) (图3.11)。5-HT在戒断后40min出现明显增高,增加幅度约为催促前的2倍(P <0.05) (图3.13)。与吗啡处理前相比,代谢产物DOPAC和5-HIAA均出现不同程度的增加(P<0.05) (图3.10,图3.12)。这些结果说明:慢性吗啡作用后给予纳络酮催促戒断可明显刺激SH-SY5Y细胞内单胺类神经递质的释放,在给予纳络酮20~40min时培养上清中单胺类神经递质含量达到高峰,其后迅速下降。
为了进一步探讨α-SNAP是否参与了吗啡处理引起的递质释放变化的调节,我们在SH-SY5Y细胞模型上又检测了吗啡作用不同时间α-SNAP的表达情况。结果发现,α-SNAP mRNA在吗啡作用的1h、6h、24h均未检测到明显的表达变化(图4.2),Western blot检测吗啡作用1h、8h、24hα-SNAP的蛋白表达亦未发现明显改变(图4.3),上述结果与动物模型上结果相一致。以上结果提示:吗啡作用不引起α-SNAP的表达变化。那么作为一个与神经递质释放关系密切的因子,它是否参与了吗啡处理引起的突触前可塑性的改变呢?
研究发现,神经递质的胞吐过程有一系列分子事件的参与,包括囊泡移动、搭靠、融合、内吞等。现在公认的递质释放中膜融合的模式是“SNARE假说”。“SNARE假说”认为,在膜融合过程中,20S复合体的聚合和解聚是其中的关键步骤。复合体的聚合始于v-SNAREs和t-SNAREs的结合。v-SNAREs和t-SNAREs分别定位于囊泡膜和质膜上,t-SNAREs包括SNAP-25和Syntaxin。膜融合发生前,t-SNAREs与v-SNAREs结合形成一个沉降系数为7S的SNARE复合物。一个7S SNARE复合物可以吸引3个α-SNAP分子,然后招募NSF使其连接至膜上。这样,SNARE、NSF和α-SNAP就形成了一个20S复合体。20S复合体一旦形成,随即在NSF的ATP酶作用下水解,同时引起SNARE分子的变构,阻止其继续聚合。SANRE解体后,形成20S复合体的各个组分又被释放到胞浆中,参与下一轮的囊泡循环。由于膜融合过程中形成20S复合体的t-SNARE位于突触前膜,所以我们推测α-SNAP在参与膜融合发生的过程中其细胞内分布有可能发生了变化,如由胞浆分布转移至胞膜。因此,我们在SH-SY5Y细胞模型上采用免疫细胞化学结合激光共聚焦检测技术又继续观察了α-SNAP在吗啡处理后的亚细胞定位。结果发现,在吗啡处理24h后给予纳络酮催促戒断20min可观测到α-SNAP开始由胞浆向胞膜转位,在随后的40min内该现象一直持续存在。在纳络酮处理60min时,转位现象仍较明显(图4.5)。而急性吗啡作用1h内未检测到上述变化(图4.4)。以上结果说明:慢性吗啡处理后给予纳络酮催促戒断可引起α-SNAP由胞浆向胞膜的转位,其转位发生的时间与递质释放变化的时间大致吻合,提示其有可能与吗啡作用引起的神经递质释放变化有关。
本研究通过测定吗啡处理及戒断不同状态下SNAPs表达、亚细胞定位及神经递质的释放情况,得出以下结论:(1)慢性吗啡处理及自然戒断对大鼠脑区NAc、CPu和Hip内α-SNAP的表达无明显影响;(2)慢性吗啡处理可上调大鼠CPu内γ-SNAP的表达,但自然戒断无上述改变,慢性吗啡处理及自然戒断对大鼠NAc和Hip内γ-SNAP的表达均无明显影响;(3)SH-SY5Y细胞可用于建立慢性吗啡依赖模型;(4)急性吗啡处理可抑制SH-SY5Y细胞单胺类神经递质的释放,慢性吗啡处理后给予纳络酮催促戒断可刺激SH-SY5Y细胞单胺类神经递质的释放;(5)急慢性吗啡处理不影响SH-SY5Y细胞内α-SNAP的表达;急性吗啡处理不引起α-SNAP亚细胞定位的改变;慢性吗啡处理后给予纳络酮催促戒断可引起α-SNAP由胞浆至胞膜的亚细胞定位发生改变,其转位可能与戒断过程中神经递质释放增加有关。
Opioid dependence is a chronic relapsing disorder in the brain characterized by compulsive drug taking, inability to limit the intake of drugs, and the emergence of withdrawal syndrome after cessation of drug taking. Nevertheless, the exact mechanisms underlying opioid dependence are still unknown. Accumulating evidence suggests that synaptic plasticity plays an important role in opioid dependence. These changes include presynaptic neurotransmitter release and postsynaptic receptor signal transduction. Previous studies demonstrated that morphine could affect the synaptic transmission by regulating molecules related with neurotransmitter release in presynapse.
Membrane fusion between synaptic vesicle membrane and presynaptic membrane has been generally received as an important step in the process of neurotransmitter release. NSF attachment proteins (SNAPs) play important roles in membrane fusion by mediating the fusion between synaptic vesicle membrane and presynaptic membrane. Three isoforms (α-,β-, andγ-) of SNAP are expressed in mammals. In view of its function in mediating membrane fusion, SNAPs take part in many important physiological processes, including regulating Ca2+-dependent exocytosis, stimulating insulin secretion in pancreatic islandβcells, increasing surfactant secretion in alveolar type II cells, participating in the nuclear envelope formation, etc.
Although considerable evidence suggests that SNAPs play key roles in the process of neurotransmitter release by mediating membrane fusion between plasma membrane and vesicle membrane, whether SNAPs are related to the presynaptic plasticity underlying opioid dependence has been not reported. To reveal the relationship between SNAPs and opioid dependence, we investigated the expression of SNAPs in different brain regions of rats with chronic morphine exposure and withdrawal, including nucleus accumbens (NAc), caudate putamen (CPu) and hippocampus (Hip), which were recognized to have close relationship with synaptic plasticity. Adult male Wistar rats were randomly assigned into control, morphine and spontaneous withdrawal group. The morphine dependent rat model was established by subcutaneous morphine injection with increasing doses for 8d, three times a day (8:00, 12:00, and 18:00). The control group was injected with the same volume of normal saline. Rats in morphine group were killed 4h after the last injection, rats in withdrawal group were killed at indicated time. Control groups were paralleled with all treatment groups. The NAc, CPu and Hip were isolated. The expression of SNAPs mRNA and protein were determined by RT-PCR and Western blot, respectively. Results showed that the expression ofγ-SNAP in CPu of morphine dependent rats was up-regulated about 25%(P<0.01) (Fig.1.4, Fig.1.9), and no alteration ofγ-SNAP in NAc and Hip was detected in morphine dependent and withdrawal groups(Fig.1.8, Fig.1.10, Table1.1, Table1.3).α-SNAP andβ-SNAP were unchanged in NAc, CPu and Hip of rats in morphine dependent and withdrawal groups. From these results above, we could draw a conclusion that morphine dependence leads to up-regulation ofγ-SNAP in CPu, but it did not have any effects on the expression ofα-SNAP andβ-SNAP in NAc, CPu and Hip(Fig1.2,Fig1.3,Fig1.5,Fig1.6, Fig1.7, Table1.1,Table1.2), which suggested that 3 isoforms of SNAPs functioned differently in morphine dependence and might be related with the secretion of specific neurotransmitter. The release of presynaptic neurotransmitter in morphine dependence and withdrawal rat was not mediated by the content ofα-SNAP andβ-SNAP, and the intracellular translocation and intrinsic activity ofα-SNAP andβ-SNAP may contribute to the molecular mechanisms underlying morphine dependence and withdrawal.
To further confirm the relationship between SNAPs and opioid dependence,α-SNAP, an isoform considered to play a key role in neurotransmitter release, was selected in the following study. Accumulating evidence reported the function ofα-SNAP in membrane fusion between vesicle membrane and presynaptic membrane, which has been regarded as the most important step in calcium-dependent exocytosis during the process of neurotransmitter release. It was reported thatα-SNAP functioned in stimulating NSF ATPase, and the effect ofα-SNAP significantly surpassed that ofγ-SNAP. Therefore, although no alteration ofα-SNAP expression was detected in animal model, this isoform was still focused on. To further study the function ofα-SNAP in morphine treatment, a well defined experimental cell model was necessarily needed owing to the relative simple and direct regulation mechanisms. The experimental model must be characterized by following traits: (1) expressing opioid receptor (2) processing similar features to mature neuroendocrine cell and secreting neurotransmitters related to opioid dependence, such as monoamine transmitters, glutamate, GABA, etc. After preliminary screening, the differentiated human neuroblastoma SH-SY5Y cell line was singled out to be applied for this study. After differentiation with retinoic acid(RA), SH-SY5Y cells were reported to show many characteristics of mature neurons, such as the appearance of extensive neurite outgrowth and electrical excitability, enhancement of neuron specific enolase(NSE) activity and increase of neurosecretory granules, etc.
In this study, we observed the formation of long neuritic processes in SH-SY5Y cells 6 days after treated with RA (Fig.2.1). The expression of NSE was also measured by Western blot. Results indicated that NSE andμopioid receptor(MOR) were up-regulated in differentiated SH-SY5Y cells compared with that in undifferentiated cells (Fig.2.2 , Fig.2.3), which was in accordance with previous research. All these properties of differentiated SH-SY5Y cells facilitated research in the molecular mechanisms of opioid dependence, especially in the aspect of neurotransmitter release related with synaptic plasticity. To elucidate the molecular mechanisms of opioid dependence, a cell model for opioid dependence was established. The cAMP overshoot after morphine withdrawal was generally applied to evaluate whether the cell model was successful. In our study, we assessed the content of intracellular cAMP by the LANCE cAMP assay and discovered cAMP overshoot in SH-SY5Y cells after naloxone-precipitated withdrawal. The content of cAMP markedly increased to a level as 2.36 fold as that in untreated cells (Fig.2.5). This confirmed that the cell model of opioid dependence in our study was successfully established.
In order to confirm the neurotransmitter changes, the contents of monoamine neurotransmitters and their metabolites were investigated in SH-SY5Y cell culture supernatant by HPLC-ECD under acute morphine treatment and naloxone-precipitated withdrawal after long-term morphine treatment. Data showed that the monoamine neurotransmitters including NE, DA and 5-HT were generally inhibited with acute morphine administration (Table3.1). After incubation with 100μM morphine for 10 min ~ 1h in SH-SY5Y cells, NE, DA and 5-HT as well as their metabolites DOPAC (DA metabolite) and 5-HIAA (5-HT metabolite) were significantly reduced, the content of NE was significantly decresed from 16.97 ng/g to 5.34 ng/g protein at 10min after morphine exposure (P<0.01). In the following 50 min, it did not significantly decline compared to that in 10min after morphine treatment and kept at a relatively low level (Fig.3.4). The changes in DA showed a similar trend to NE , that was, an obvious decline was detected in the first 10 min, lower than the basal level (P<0.01)(Fig.3.6). The content of 5-HT was significantly decreased (P<0.05) in the initial 10 min and continuted decline till 1/3 of the original level at 1h after morphine treatment (P<0.01) (Fig.3.8). Their metabolites, DOPAC and 5-HIAA were also down-regulated compared with cells without exposure to morphine (P<0.01) (Fig.3.5, Fig.3.7). These results showed that acute morphine treatment significantly attenuated the release of monoamine neurotransmitters.
When exposed to 100μM naloxone after incubation of morphine for 24h, monoamine neurotransmitter release was investigated in SH-SY5Y cell supernatants. Results showed that NE, DA and 5-HT and their metabolites, DOPAC and 5-HIAA increased (Table3.2). The content of NE rose from 7.59 ng/g to 12.74 ng/g, higher nearly 0.5 fold, at 20 min after the onset of withdrawal(P<0.01), then dropped to the basal level at 40 min after the onset of withdrawal (Fig.3.9). Compared with NE, the change in DA was more obvious, and the DA content reached a peak at 20 min after withdrawal, approximately twice the original level(P<0.05)(Fig.3.11). 5-HT was also detected a significant increase at 40 min, nearly twice the level of untreated ones (P<0.05) (Fig.3.13). Their metabolites, DOPAC and 5-HIAA, also increased, compared with that of cells before naloxone treatment(P<0.05) (Fig.3.10, Fig.3.12). These results showed that naloxone-precipitated withdrawal after morphine dependence significantly stimulated the release of monoamine neurotransmitters. The content of monoamine neurotransmitters reached peaks at 20~40min after the onset of naloxone withdrawal, then dropped rapidly in the following time.
To determine whetherα-SNAP participated in the changes of neurotransmitters induced by morphine, theα-SNAP expression was detected in SH-SY5Y cells after morphine administration. Data showed that no alteration ofα-SNAP mRNA expression was detected at 1h, 6h, 24h in SH-SY5Y cells after morphine administration (Fig.4.2), and there was no change inα-SNAP protein expression at 1h, 8h, 24h after morphine treatment by Western blot analysis (Fig.4.3). The results obtained from cells were in accordance with that obtained from animal model. Data suggested that morphine treatment did not result in any change inα-SNAP expression. However, as a key role in neurotransmitter release, whetherα-SNAP takes part in presynaptic plasticity underlying morphine dependence has not been elucidated.
Many evidences showed that a series of molecular events take place in the process of neurotransmitter exocytosis, including vesicle trafficking, docking, fusion and internalization.“SNARE hypothesis”is widely accepted in membrane fusion during the neurotransmitter release. According to“SNARE hypothesis”, in the process of membrane fusion, 20S complex assembly and disassembly are the key steps. Complex assembly begins with binding between v-SNAREs and t-SNAREs. v-SNAREs and t-SNAREs are located in vesicle membrane and target membrane, respectively. t-SNAREs include SNAP-25 and Syntaxin. At the beginning of membrane fusion, t-SNAREs bound to v-SNAREs, and 7S SNARE complex is formed. One 7S SNARE complex can recruit 3α-SNAP molecules, this is followed by the binding of NSF. Thus, SNAREs,α-SNAP and NSF assembled a 20S complex. Then, subsequent ATP hydrolysis of NSF leads to disassembly of 20S complex and a conformational change in SNARE molecules, which prevented from its reassembly. Thus, all components of SNAREs enter into next cycle of vesicle transport. Since t-SNARE, a key element in the formation of 20S complex, located at presynaptic membrane, we supposed that the translocation ofα-SNAP might take place in the process of membrane fusion, for instance, from cytoplasm to cell membrane. Therefore, we further examined the subcellular location ofα-SNAP by confocal laser scanning microscopy in SH-SY5Y cells with morphine treatment. Results showed thatα-SNAP began to migrate from cytoplasm to cell membrane at 20min after naloxone withdrawal and lasted in the following 40 minutes. An obvious translocation was still observed at 60min after withdrawal (Fig.4.5). Nevertheless, no change was detected in subcellular location ofα-SNAP with acute morphine treatment (Fig.4.4). It showed that naloxone-precipitated withdrawal after chronic morphine treatment altered the subcellular localization ofα-SNAP in SH-SY5Y cells, and the time courses of changes in monoamine neurotransmitters approximately coincided with the time ofα-SNAP translocation, which might indicate a close relationship between neurotransmitter release andα-SNAP translocation.
In this study, we drew the following conclusions through determination of SNAPs expression and subcellular location as well as neurotransmitters release after morphine treatment: (1) Chronic morphine administration and spontaneous withdrawal did not affectα-SNAP expression in NAc, CPu and Hip of rats. (2) Chronic morphine administration up-regulatedγ-SNAP expression in CPu of rats, but did not changeγ-SNAP expression in NAc and Hip of rats. Spontaneous withdrawal had no effect onγ-SNAP expression in NAc, CPu and Hip. (3) SH-SY5Y cells could be used to establish cell model of morphine dependence. (4) Acute morphine treatment significantly attenuated the release of monoamine neurotransmitters, while naloxone-precipitated withdrawal significantly stimulated the release of monoamine neurotransmitters. (5) Acute and chronic morphine treatment did not affectα-SNAP expression in SH-SY5Y cells. Acute morphine treatment did not alter the subcellular localization ofα-SNAP in SH-SY5Y cells. Naloxone-precipitated withdrawal after morphine dependence resulted inα-SNAP translocation from cytoplasm to cell membrane, which might be related with the neurotransmitter release after morphine withdrawal.
在神经递质的释放过程中,囊泡膜和质膜的融合是影响递质释放的关键步骤。可溶性NSF附着蛋白(Soluble NSF attachment proteins,SNAPs)是一类在膜融合过程中发挥重要作用的蛋白质分子,介导了囊泡膜和质膜的融合,在哺乳动物中存在α-,β-,和γ- 3种亚型。鉴于SNAPs在膜融合过程中发挥的重要作用,SNAPs参与许多重要的生理进程,包括调节钙依赖的胞吐机制、刺激胰岛β细胞分泌胰岛素、增加肺泡II型细胞表面活性物质的分泌、参与有丝分裂过程中核膜的形成等。
虽然大量研究表明SNAPs在神经递质释放的膜融合过程中起重要作用,但是SNAPs是否参与了阿片依赖过程中突触前神经可塑性的发生目前尚未见报道。为了揭示SNAPs与阿片依赖的关系,我们首先在大鼠吗啡依赖及戒断模型上观察了与神经可塑性关系密切的脑区伏隔核(nucleus accumbens,NAc)、尾壳核(caudate putamen,CPu)及海马(hippocampus,Hip)中SNAPs表达的变化,初步探讨SNAPs和吗啡依赖的关系。研究采用成年雄性Wistar大鼠,随机分为对照组、吗啡组和戒断不同时间组。吗啡组大鼠背部皮下注射吗啡8d,每日3次,剂量递增,对照组大鼠注射等体积生理盐水。吗啡组末次注射吗啡4 h后处死,断头取脑。自然戒断组分别在戒断不同时间处死动物。各组均设平行对照。利用RT-PCR和Western blot技术分别检测各脑区SNAPs的mRNA和蛋白表达水平。结果发现,与对照组相比,慢性吗啡依赖大鼠CPu内γ-SNAP的mRNA和蛋白表达水平均上调25%左右(P<0.01) (图1.4,图1.9),NAc及Hip脑区则无明显变化,自然戒断d2、d3、d7组亦未观察到γ-SNAP明显的表达改变(图1.8,图1.10,表1.1,表1.3)。α-SNAP和β-SNAP在吗啡依赖和自然戒断状态下3个被检脑区(NAc、CPu、Hip)均未检测到明显的表达变化(图1.2,图1.3,图1.5,图1.6,图1.7,表1.1,表1.2)。以上结果说明:慢性吗啡依赖可增加CPu内γ-SNAP的表达,对α-SNAP和β-SNAP的表达没有影响,提示SNAPs 3种亚型在慢性吗啡依赖过程中行使不同的功能,可能与特定神经递质的分泌有关。慢性吗啡依赖及戒断引起的突触前神经递质释放改变可能不是由α-SNAP和β-SNAP表达量变化介导的,其具体机制可能与其内在活性变化或蛋白在细胞内的分布有关。
为了进一步研究SNAPs与阿片依赖的关系,我们选择了一种与神经递质释放关系最为明确的亚型—α-SNAP进行研究。已有大量研究报道α-SNAP参与钙依赖的胞吐作用,该作用在神经递质释放过程中的囊泡膜与质膜的融合步骤中是至关重要的。且相关研究报道α-SNAP通过刺激NSF的ATP酶活性来发挥其在递质释放中的作用,α-SNAP对NSF的ATP酶刺激作用明显强于γ-SNAP。因此,虽然我们在动物模型上未检测到α-SNAP的蛋白表达变化,但由于其在递质释放中的重要作用,我们依然选择这种亚型作为研究对象。为了进一步研究α-SNAP的功能与吗啡依赖的关系,选择一个良好的、调控模式相对简单和直接的细胞模型是十分必要的。该细胞模型必须能够满足以下条件:(1)表达阿片受体;(2)拥有类似成熟神经内分泌细胞的一些特性,能释放与阿片依赖有关的神经递质,如单胺类、谷氨酸(glutamate,Glu)、γ-氨基丁酸(γ-aminobutyric acid ,GABA)等。经过初步筛选,我们选择了分化的人神经母细胞瘤细胞株SH-SY5Y作为研究对象。据研究报道,SH-SY5Y细胞株经过维甲酸(retinoic acid,RA)分化后可表现出许多类似成熟神经元的特性,如神经突起变长、电刺激的兴奋性增加、神经元特异性烯醇化酶(neuron specific enolase,NSE)表达增加、神经分泌颗粒增多等。
在研究中我们发现,RA分化6天的SH-SY5Y细胞可形成较长的神经突起(图2.1),免疫印迹分析检测到分化后的细胞内NSE表达增加(图2.2)。另外,在我们的研究中亦发现了RA分化后的细胞μ阿片受体(μopioid receptor,MOR)表达上调,与以往研究相一致(图2.3)。这些特性都为我们研究阿片依赖的分子机制提供了有力的条件,尤其是与突触前神经递质释放有关的分子机制。为了研究与阿片依赖有关的分子机制,我们首先需要在SH-SY5Y细胞上建立一个阿片依赖的细胞模型。阿片长期暴露后给予纳络酮催促戒断出现cAMP超射常常被作为评定阿片依赖模型建立是否成功的一个重要指标。本研究采用免疫竞争结合法,测定了吗啡处理前后细胞内cAMP含量的变化,发现100μM吗啡作用分化SH-SY5Y细胞24h后给予纳络酮催促戒断可形成明显的cAMP超射,cAMP含量约为催促前的2.36倍(图2.5),证实了分化SH-SY5Y细胞上吗啡依赖的细胞模型建立是成功的。
为了进一步确定吗啡处理过程中神经递质的释放情况,我们采用高效液相-电化学检测法(high performance liquid chromatography- electrochemical detection,HPLC-ECD)测定了急性吗啡作用及慢性吗啡作用纳络酮催促戒断后不同时间SH-SY5Y细胞培养上清液中单胺类神经递质的含量,探讨吗啡处理与神经递质释放的关系。结果显示,在急性吗啡作用下,单胺类神经递质,包括去甲肾上腺素(norepinephrine,NE)、多巴胺(dopamine,DA)、5-羟色胺(5-hydroxytryptamine,5-HT)的释放均受到普遍抑制(表3.1)。100μM吗啡孵育SH-SY5Y细胞10min~1h,细胞培养上清内NE,DA和5-HT及其代谢产物3,4-二羟基苯乙酸(3,4-dihydroxy- phenylacetic acid,DOPAC) (DA代谢产物)和5-羟基吲哚乙酸(5-hydroxyindolacetic acid,5-HIAA )(5-HT代谢产物)均显著下降,其中NE含量从16.97 ng/g蛋白下降至5.34 ng/g蛋白,下降幅度以吗啡作用10min内下降最为明显(P <0.01),在吗啡继续作用的50min内一直保持在较低水平(图3.4)。DA的变化趋势与NE类似,在吗啡作用的最初10min亦出现明显下降,而且下降幅度在2/3以上(P <0.01) (图3.6)。虽然5-HT在吗啡急性作用的早期出现中等程度的下降,但在吗啡继续作用的50min内,5-HT含量继续降低,至吗啡作用后1h达到基础水平的1/3左右(P<0.01)(图3.8)。与吗啡处理前相比,代谢产物DOPAC和5-HIAA均出现不同程度的降低(P<0.01) (图3.5,图3.7)。这些结果表明,急性吗啡作用可以显著抑制SH-SY5Y细胞单胺类神经递质的释放。
在给予100μM吗啡孵育细胞24h后用100μM纳络酮催促戒断测定SH-SY5Y细胞上清单胺类神经递质的含量,结果发现:NE、DA和5-HT及其代谢产物DOPAC和5-HIAA均有不同程度的增加(表3.2)。NE的含量在戒断20 min后由戒断前7.59 ng/g蛋白增加至12.74 ng/g蛋白,增加50%左右,与催促前相比有显著性差异(P <0.01),在戒断40min时恢复至戒断前水平(图3.9)。与NE相比,DA的变化趋势更加明显,DA在戒断的10min即有增加趋势,但与催促前相比无显著性差异(P> 0.05),在戒断20min时达最高值,约为催促前基础水平的2倍(P <0.05) (图3.11)。5-HT在戒断后40min出现明显增高,增加幅度约为催促前的2倍(P <0.05) (图3.13)。与吗啡处理前相比,代谢产物DOPAC和5-HIAA均出现不同程度的增加(P<0.05) (图3.10,图3.12)。这些结果说明:慢性吗啡作用后给予纳络酮催促戒断可明显刺激SH-SY5Y细胞内单胺类神经递质的释放,在给予纳络酮20~40min时培养上清中单胺类神经递质含量达到高峰,其后迅速下降。
为了进一步探讨α-SNAP是否参与了吗啡处理引起的递质释放变化的调节,我们在SH-SY5Y细胞模型上又检测了吗啡作用不同时间α-SNAP的表达情况。结果发现,α-SNAP mRNA在吗啡作用的1h、6h、24h均未检测到明显的表达变化(图4.2),Western blot检测吗啡作用1h、8h、24hα-SNAP的蛋白表达亦未发现明显改变(图4.3),上述结果与动物模型上结果相一致。以上结果提示:吗啡作用不引起α-SNAP的表达变化。那么作为一个与神经递质释放关系密切的因子,它是否参与了吗啡处理引起的突触前可塑性的改变呢?
研究发现,神经递质的胞吐过程有一系列分子事件的参与,包括囊泡移动、搭靠、融合、内吞等。现在公认的递质释放中膜融合的模式是“SNARE假说”。“SNARE假说”认为,在膜融合过程中,20S复合体的聚合和解聚是其中的关键步骤。复合体的聚合始于v-SNAREs和t-SNAREs的结合。v-SNAREs和t-SNAREs分别定位于囊泡膜和质膜上,t-SNAREs包括SNAP-25和Syntaxin。膜融合发生前,t-SNAREs与v-SNAREs结合形成一个沉降系数为7S的SNARE复合物。一个7S SNARE复合物可以吸引3个α-SNAP分子,然后招募NSF使其连接至膜上。这样,SNARE、NSF和α-SNAP就形成了一个20S复合体。20S复合体一旦形成,随即在NSF的ATP酶作用下水解,同时引起SNARE分子的变构,阻止其继续聚合。SANRE解体后,形成20S复合体的各个组分又被释放到胞浆中,参与下一轮的囊泡循环。由于膜融合过程中形成20S复合体的t-SNARE位于突触前膜,所以我们推测α-SNAP在参与膜融合发生的过程中其细胞内分布有可能发生了变化,如由胞浆分布转移至胞膜。因此,我们在SH-SY5Y细胞模型上采用免疫细胞化学结合激光共聚焦检测技术又继续观察了α-SNAP在吗啡处理后的亚细胞定位。结果发现,在吗啡处理24h后给予纳络酮催促戒断20min可观测到α-SNAP开始由胞浆向胞膜转位,在随后的40min内该现象一直持续存在。在纳络酮处理60min时,转位现象仍较明显(图4.5)。而急性吗啡作用1h内未检测到上述变化(图4.4)。以上结果说明:慢性吗啡处理后给予纳络酮催促戒断可引起α-SNAP由胞浆向胞膜的转位,其转位发生的时间与递质释放变化的时间大致吻合,提示其有可能与吗啡作用引起的神经递质释放变化有关。
本研究通过测定吗啡处理及戒断不同状态下SNAPs表达、亚细胞定位及神经递质的释放情况,得出以下结论:(1)慢性吗啡处理及自然戒断对大鼠脑区NAc、CPu和Hip内α-SNAP的表达无明显影响;(2)慢性吗啡处理可上调大鼠CPu内γ-SNAP的表达,但自然戒断无上述改变,慢性吗啡处理及自然戒断对大鼠NAc和Hip内γ-SNAP的表达均无明显影响;(3)SH-SY5Y细胞可用于建立慢性吗啡依赖模型;(4)急性吗啡处理可抑制SH-SY5Y细胞单胺类神经递质的释放,慢性吗啡处理后给予纳络酮催促戒断可刺激SH-SY5Y细胞单胺类神经递质的释放;(5)急慢性吗啡处理不影响SH-SY5Y细胞内α-SNAP的表达;急性吗啡处理不引起α-SNAP亚细胞定位的改变;慢性吗啡处理后给予纳络酮催促戒断可引起α-SNAP由胞浆至胞膜的亚细胞定位发生改变,其转位可能与戒断过程中神经递质释放增加有关。
Opioid dependence is a chronic relapsing disorder in the brain characterized by compulsive drug taking, inability to limit the intake of drugs, and the emergence of withdrawal syndrome after cessation of drug taking. Nevertheless, the exact mechanisms underlying opioid dependence are still unknown. Accumulating evidence suggests that synaptic plasticity plays an important role in opioid dependence. These changes include presynaptic neurotransmitter release and postsynaptic receptor signal transduction. Previous studies demonstrated that morphine could affect the synaptic transmission by regulating molecules related with neurotransmitter release in presynapse.
Membrane fusion between synaptic vesicle membrane and presynaptic membrane has been generally received as an important step in the process of neurotransmitter release. NSF attachment proteins (SNAPs) play important roles in membrane fusion by mediating the fusion between synaptic vesicle membrane and presynaptic membrane. Three isoforms (α-,β-, andγ-) of SNAP are expressed in mammals. In view of its function in mediating membrane fusion, SNAPs take part in many important physiological processes, including regulating Ca2+-dependent exocytosis, stimulating insulin secretion in pancreatic islandβcells, increasing surfactant secretion in alveolar type II cells, participating in the nuclear envelope formation, etc.
Although considerable evidence suggests that SNAPs play key roles in the process of neurotransmitter release by mediating membrane fusion between plasma membrane and vesicle membrane, whether SNAPs are related to the presynaptic plasticity underlying opioid dependence has been not reported. To reveal the relationship between SNAPs and opioid dependence, we investigated the expression of SNAPs in different brain regions of rats with chronic morphine exposure and withdrawal, including nucleus accumbens (NAc), caudate putamen (CPu) and hippocampus (Hip), which were recognized to have close relationship with synaptic plasticity. Adult male Wistar rats were randomly assigned into control, morphine and spontaneous withdrawal group. The morphine dependent rat model was established by subcutaneous morphine injection with increasing doses for 8d, three times a day (8:00, 12:00, and 18:00). The control group was injected with the same volume of normal saline. Rats in morphine group were killed 4h after the last injection, rats in withdrawal group were killed at indicated time. Control groups were paralleled with all treatment groups. The NAc, CPu and Hip were isolated. The expression of SNAPs mRNA and protein were determined by RT-PCR and Western blot, respectively. Results showed that the expression ofγ-SNAP in CPu of morphine dependent rats was up-regulated about 25%(P<0.01) (Fig.1.4, Fig.1.9), and no alteration ofγ-SNAP in NAc and Hip was detected in morphine dependent and withdrawal groups(Fig.1.8, Fig.1.10, Table1.1, Table1.3).α-SNAP andβ-SNAP were unchanged in NAc, CPu and Hip of rats in morphine dependent and withdrawal groups. From these results above, we could draw a conclusion that morphine dependence leads to up-regulation ofγ-SNAP in CPu, but it did not have any effects on the expression ofα-SNAP andβ-SNAP in NAc, CPu and Hip(Fig1.2,Fig1.3,Fig1.5,Fig1.6, Fig1.7, Table1.1,Table1.2), which suggested that 3 isoforms of SNAPs functioned differently in morphine dependence and might be related with the secretion of specific neurotransmitter. The release of presynaptic neurotransmitter in morphine dependence and withdrawal rat was not mediated by the content ofα-SNAP andβ-SNAP, and the intracellular translocation and intrinsic activity ofα-SNAP andβ-SNAP may contribute to the molecular mechanisms underlying morphine dependence and withdrawal.
To further confirm the relationship between SNAPs and opioid dependence,α-SNAP, an isoform considered to play a key role in neurotransmitter release, was selected in the following study. Accumulating evidence reported the function ofα-SNAP in membrane fusion between vesicle membrane and presynaptic membrane, which has been regarded as the most important step in calcium-dependent exocytosis during the process of neurotransmitter release. It was reported thatα-SNAP functioned in stimulating NSF ATPase, and the effect ofα-SNAP significantly surpassed that ofγ-SNAP. Therefore, although no alteration ofα-SNAP expression was detected in animal model, this isoform was still focused on. To further study the function ofα-SNAP in morphine treatment, a well defined experimental cell model was necessarily needed owing to the relative simple and direct regulation mechanisms. The experimental model must be characterized by following traits: (1) expressing opioid receptor (2) processing similar features to mature neuroendocrine cell and secreting neurotransmitters related to opioid dependence, such as monoamine transmitters, glutamate, GABA, etc. After preliminary screening, the differentiated human neuroblastoma SH-SY5Y cell line was singled out to be applied for this study. After differentiation with retinoic acid(RA), SH-SY5Y cells were reported to show many characteristics of mature neurons, such as the appearance of extensive neurite outgrowth and electrical excitability, enhancement of neuron specific enolase(NSE) activity and increase of neurosecretory granules, etc.
In this study, we observed the formation of long neuritic processes in SH-SY5Y cells 6 days after treated with RA (Fig.2.1). The expression of NSE was also measured by Western blot. Results indicated that NSE andμopioid receptor(MOR) were up-regulated in differentiated SH-SY5Y cells compared with that in undifferentiated cells (Fig.2.2 , Fig.2.3), which was in accordance with previous research. All these properties of differentiated SH-SY5Y cells facilitated research in the molecular mechanisms of opioid dependence, especially in the aspect of neurotransmitter release related with synaptic plasticity. To elucidate the molecular mechanisms of opioid dependence, a cell model for opioid dependence was established. The cAMP overshoot after morphine withdrawal was generally applied to evaluate whether the cell model was successful. In our study, we assessed the content of intracellular cAMP by the LANCE cAMP assay and discovered cAMP overshoot in SH-SY5Y cells after naloxone-precipitated withdrawal. The content of cAMP markedly increased to a level as 2.36 fold as that in untreated cells (Fig.2.5). This confirmed that the cell model of opioid dependence in our study was successfully established.
In order to confirm the neurotransmitter changes, the contents of monoamine neurotransmitters and their metabolites were investigated in SH-SY5Y cell culture supernatant by HPLC-ECD under acute morphine treatment and naloxone-precipitated withdrawal after long-term morphine treatment. Data showed that the monoamine neurotransmitters including NE, DA and 5-HT were generally inhibited with acute morphine administration (Table3.1). After incubation with 100μM morphine for 10 min ~ 1h in SH-SY5Y cells, NE, DA and 5-HT as well as their metabolites DOPAC (DA metabolite) and 5-HIAA (5-HT metabolite) were significantly reduced, the content of NE was significantly decresed from 16.97 ng/g to 5.34 ng/g protein at 10min after morphine exposure (P<0.01). In the following 50 min, it did not significantly decline compared to that in 10min after morphine treatment and kept at a relatively low level (Fig.3.4). The changes in DA showed a similar trend to NE , that was, an obvious decline was detected in the first 10 min, lower than the basal level (P<0.01)(Fig.3.6). The content of 5-HT was significantly decreased (P<0.05) in the initial 10 min and continuted decline till 1/3 of the original level at 1h after morphine treatment (P<0.01) (Fig.3.8). Their metabolites, DOPAC and 5-HIAA were also down-regulated compared with cells without exposure to morphine (P<0.01) (Fig.3.5, Fig.3.7). These results showed that acute morphine treatment significantly attenuated the release of monoamine neurotransmitters.
When exposed to 100μM naloxone after incubation of morphine for 24h, monoamine neurotransmitter release was investigated in SH-SY5Y cell supernatants. Results showed that NE, DA and 5-HT and their metabolites, DOPAC and 5-HIAA increased (Table3.2). The content of NE rose from 7.59 ng/g to 12.74 ng/g, higher nearly 0.5 fold, at 20 min after the onset of withdrawal(P<0.01), then dropped to the basal level at 40 min after the onset of withdrawal (Fig.3.9). Compared with NE, the change in DA was more obvious, and the DA content reached a peak at 20 min after withdrawal, approximately twice the original level(P<0.05)(Fig.3.11). 5-HT was also detected a significant increase at 40 min, nearly twice the level of untreated ones (P<0.05) (Fig.3.13). Their metabolites, DOPAC and 5-HIAA, also increased, compared with that of cells before naloxone treatment(P<0.05) (Fig.3.10, Fig.3.12). These results showed that naloxone-precipitated withdrawal after morphine dependence significantly stimulated the release of monoamine neurotransmitters. The content of monoamine neurotransmitters reached peaks at 20~40min after the onset of naloxone withdrawal, then dropped rapidly in the following time.
To determine whetherα-SNAP participated in the changes of neurotransmitters induced by morphine, theα-SNAP expression was detected in SH-SY5Y cells after morphine administration. Data showed that no alteration ofα-SNAP mRNA expression was detected at 1h, 6h, 24h in SH-SY5Y cells after morphine administration (Fig.4.2), and there was no change inα-SNAP protein expression at 1h, 8h, 24h after morphine treatment by Western blot analysis (Fig.4.3). The results obtained from cells were in accordance with that obtained from animal model. Data suggested that morphine treatment did not result in any change inα-SNAP expression. However, as a key role in neurotransmitter release, whetherα-SNAP takes part in presynaptic plasticity underlying morphine dependence has not been elucidated.
Many evidences showed that a series of molecular events take place in the process of neurotransmitter exocytosis, including vesicle trafficking, docking, fusion and internalization.“SNARE hypothesis”is widely accepted in membrane fusion during the neurotransmitter release. According to“SNARE hypothesis”, in the process of membrane fusion, 20S complex assembly and disassembly are the key steps. Complex assembly begins with binding between v-SNAREs and t-SNAREs. v-SNAREs and t-SNAREs are located in vesicle membrane and target membrane, respectively. t-SNAREs include SNAP-25 and Syntaxin. At the beginning of membrane fusion, t-SNAREs bound to v-SNAREs, and 7S SNARE complex is formed. One 7S SNARE complex can recruit 3α-SNAP molecules, this is followed by the binding of NSF. Thus, SNAREs,α-SNAP and NSF assembled a 20S complex. Then, subsequent ATP hydrolysis of NSF leads to disassembly of 20S complex and a conformational change in SNARE molecules, which prevented from its reassembly. Thus, all components of SNAREs enter into next cycle of vesicle transport. Since t-SNARE, a key element in the formation of 20S complex, located at presynaptic membrane, we supposed that the translocation ofα-SNAP might take place in the process of membrane fusion, for instance, from cytoplasm to cell membrane. Therefore, we further examined the subcellular location ofα-SNAP by confocal laser scanning microscopy in SH-SY5Y cells with morphine treatment. Results showed thatα-SNAP began to migrate from cytoplasm to cell membrane at 20min after naloxone withdrawal and lasted in the following 40 minutes. An obvious translocation was still observed at 60min after withdrawal (Fig.4.5). Nevertheless, no change was detected in subcellular location ofα-SNAP with acute morphine treatment (Fig.4.4). It showed that naloxone-precipitated withdrawal after chronic morphine treatment altered the subcellular localization ofα-SNAP in SH-SY5Y cells, and the time courses of changes in monoamine neurotransmitters approximately coincided with the time ofα-SNAP translocation, which might indicate a close relationship between neurotransmitter release andα-SNAP translocation.
In this study, we drew the following conclusions through determination of SNAPs expression and subcellular location as well as neurotransmitters release after morphine treatment: (1) Chronic morphine administration and spontaneous withdrawal did not affectα-SNAP expression in NAc, CPu and Hip of rats. (2) Chronic morphine administration up-regulatedγ-SNAP expression in CPu of rats, but did not changeγ-SNAP expression in NAc and Hip of rats. Spontaneous withdrawal had no effect onγ-SNAP expression in NAc, CPu and Hip. (3) SH-SY5Y cells could be used to establish cell model of morphine dependence. (4) Acute morphine treatment significantly attenuated the release of monoamine neurotransmitters, while naloxone-precipitated withdrawal significantly stimulated the release of monoamine neurotransmitters. (5) Acute and chronic morphine treatment did not affectα-SNAP expression in SH-SY5Y cells. Acute morphine treatment did not alter the subcellular localization ofα-SNAP in SH-SY5Y cells. Naloxone-precipitated withdrawal after morphine dependence resulted inα-SNAP translocation from cytoplasm to cell membrane, which might be related with the neurotransmitter release after morphine withdrawal.
引文
1 Leshner AI. Addiction is a brain disease, and it matters. Science (New York, N.Y.), 1997, 278(5335):45-7
2 Nestler EJ, Aghajanian GK. Molecular and cellular basis of addiction. Science (New York, N.Y.), 1997, 278(5335):58-63
3 Williams JT, Christie MJ, Manzoni O. Cellular and synaptic adaptations mediating opioid dependence. Physiol Rev, 2001, 81(1):299-343
4 Taylor DA, Fleming WW. Unifying perspectives of the mechanisms underlying the development of tolerance and physical dependence to opioids. J Pharmacol Exp Ther, 2001, 297(1):11-18
5 曹君利,丁海雷,杨国栋,等.囊泡胞吐机制及与其相关的神经元可塑性.生命科学,2003,5(15): 283-288
6 蔡倩,陆佩华,盛祖抗.神经末梢突触囊泡神经递质过程的调控蛋白.生理科学进展,2003,1(34): 6-10
7 王堰婷,陆佩华,盛祖抗.神经递质释放过程中的可溶性 NSF 附着蛋白受体(SNARE)和相关蛋白.生物化学与生物物理进展,2002,29(1): 35-38
8 Sollner T, Whiteheart SW, Brunner M, et al. SNAP receptors implicated in vesicle targeting and fusion. Nature, 1993, 362(6418):318-24
9 Clary DO, Rothman JE. Purification of three related peripheral membrane proteins needed for vesicular transport. J Biol Chem, 1990,265(17): 10109-17
10 Whiteheart SW, Griff IC, Brunner M, et al. SNAP family of NSFattachment proteins includes a brain-specific isoform. Nature, 1993, 362(6418):353-5
11 Haynes LP, Barnard RJ, Morgan A, et al. Stimulation of NSF ATPase activity during t-SNARE priming. FEBS Lett, 1998, 436(1):1-5
12 Morgan A, Dimaline R, Burgoyne RD. The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. J Biol Chem, 1994, 269(47):29347-50
13 De Souza Silva MA, Mattern C, Hacker R, et al. Intranasal administration of the dopaminergic agonists L-DOPA, amphetamine, and cocaine increases dopamine activity in the neostriatum: a microdialysis study in the rat. J Neurochem, 1997, 68(1):233-9
14 Suzuki T, Tsuji M, Mori T, et al. Effect of naltrindole on the development of physical dependence on morphine in mice: a behavioral and biochemical study. Life Sci, 1995, 57(17):PL247-52
15 Wise RA, Bozarth MA. A psychomotor stimulant theory of addiction. Psychol Rev, 1987, 94(4):469-92
16 Leone P, Pocock D, Wise RA. Morphine-dopamine interaction: ventral tegmental morphine increases nucleus accumbens dopamine release. Pharmacol Biochem Behav, 1991, 39(2):469 -72
17 Di Chiara G, Imperato A. Drugs abused by humans preferentially increase synaptic dopamine concentrations in the mesolimbic system of freely moving rats. Proc Natl Acad Sci U S A, 1988, 85(14):5274-8
18 Gerra G, Zaimovic A, Moi G, et al. Aggressive responding in abstinent heroin addicts: neuroendocrine and personality correlates. Prog Neuropsychopharmacol Biol Psychiatry, 2004, 28(1):129-39
19 Miao H, Qin By, Yang Y, et al. Ultrastructural changes in rat locus coeruleus induced by chronic opioids. Acta Neuropatho, 1997;94:109
20 Kamei J, Ohsawa M. Role of noradrenergic functions in the modification of naloxone-precipitated withdrawal jumping in morphine-dependent mice by diabetes. Life Sci, 1997, 60(15):PL223 -8
21 Mylecharane EJ. Ventral tegmental area 5-HT receptors: mesolimbicdopamine release and behavioural studies. Behav Brain Res, 1996, 73 (1-2):1-5
22 方芳,曹清,宋福津,等.吗啡长时程对 SH-SY5Y 细胞 cAMP 系统及c-Fos 磷酸化的影响.中国医学科学学报,1999,4(21): 262-267
23 Xu NJ, Yu YX, Zhu JM, et al. Inhibition of SNAP - 25 phosphorylation at Ser187 is involved in chronic morphine - induced down - regulation of SNARE complex formation [J]. J Biol Chem,2004,279 (39):40601– 40608
24 Manzoni OJ, Williams JT. Presynaptic regulation of glutamate release in the ventral tegmental area during morphine withdrawal [J]. J Neurosci, 1999, 19:6629–6636
25 Sepulveda MJ, Hernandez L, Rada P, et al. Effect of precipitated withdrawal on extracellular glutamate and aspartate in the nucleus accumbens of chronically morphine-treated rats: an in vivo microdialysis study [J]. Pharmacol Biochem Behav, 1998, 60:255–262
26 Sudlow AW, McFerran BW, Bodill H, Barnard RJ, Morgan A, Burgoyne RD.Similar effects of α- and β-SNAP on Ca2+-regulated exocytosis. FEBS Lett. 1996 Sep 16;393(2-3):185~8
27 Nagamatsu S, Watanabe T, Nakamichi Y, et al. α-soluble N-ethylmaleimide-sensitive factor attachment protein is expressed in pancreatic β cells and functions in insulin but not γ-aminobutyric acid secretion. J Biol Chem, 1999, 274(12):8053-60
28 Abonyo BO, Wang P, Narasaraju TA, et al. Characterization of α-Soluble N-Ethylmaleimide–Sensitive Fusion Attachment Protein in Alveolar Type II Cells: Implications in Lung Surfactant Secretion. Am J Respir Cell Mol Biol, 2003,29:273–282
29 Baur T, Ramadan K, Schlundt A, et al. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts. J Cell Sci, 2007, 120:2895-903
30 George Paxinos,Charles Watson,主编. 诸葛启钏,主译.大鼠脑立体定位图谱[M]. 第 3 版,北京:人民卫生出版社,2005
31 DeBello WM, O'Connor V, Dresbach T, et al. SNAP-mediated protein-protein interactions essential for neurotransmitter release. Nature, 1995, 373(6515):626-30
32 Morgan A, Burgoyne RD. A role for soluble NSF attachment proteins (SNAPs) in regulated exocytosis in adrenal chromaffin cells. EMBO, 1995, 14(2):232-9
33 Xu J, Xu Y, Ellis-Davies GC,et al. Differential Regulation of Exocytosis by α- and β-SNAPs [J]. J of Neurosci,2002,22(1):53-61
34 Nakajima K, Hirose H, Taniguchi M, et al. Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion. EMBO J, 2004, 23:3216-26
35 Schiavo G, Gmachl MJ, Stenbeck G, et al. A possible docking and fusion particle for synaptic transmission. Nature, 1995, 378:733-6
36 Hanley JG, Khatri L, Hanson PI, et al. NSF ATPase and α-/β-SNAPs disassemble the AMPA receptor-PICK1 complex. Neuron, 2002, 34:53-67
37 Chen D, Xu W, He P, et al. Gaf-1, a γ -SNAP-binding protein associated with the mitochondria. J Biol Chem, 2001, 276:13127-35
38 Tani K, Shibata M, Kawase K, et al. Mapping of functional domains of γ-SNAP. J Biol Chem, 2003, 278:13531-8
39 Nakamichi Y, Nagamatsu S. α-SNAP functions in insulin exocytosis from mature, but not immature secretory granules in pancreatic β cells [J]. Biochem Biophys Res Commun,1999,260(1):127-132
1 Yu VC, Richards ML, Sadee W. A human neuroblastoma cell line expresses mu and delta opioid receptor sites. J Biol Chem, 1986,261(3):1065-70
2 Yu VC, Sadee W. Efficacy and tolerance of narcotic analgesics at the mu opioid receptor in differentiated human neuroblastoma cells. J Pharmacol Exp Ther, 1988,245(1):350-5
3 Odelstad L, Pahlman S, Nilsson K, et al. Neuron-specific enolase in relation to differentiation in human neuroblastoma. Brain Res, 1981,224(1):69-82
4 Zadina JE, Chang SL, Ge LJ, et al. Mu opiate receptor down-regulation by morphine and up-regulation by naloxone in SH-SY5Y human neuroblastoma cells. J Pharmacol Exp Ther, 1993,265(1):254-62
5 Kam AY, Chan AS, Wong YH. Phosphatidylinositol-3 kinase is distinctively required for mu-, but not kappa-opioid receptor-induced activation of c-Jun N-terminal kinase. J Neurochem,2004,89(2):391-402
6 Hou YN, Vlaskovska M, Cebers G, et al. A mu-receptor opioid agonist induces AP-1 and NF-kappa B transcription factor activity in primary cultures of rat cortical neurons. Neurosci Lett, 1996,212(3):159-62
7 Yu VC, Eiger S, Duan DS, et al. Regulation of cyclic AMP by the mu-opioid receptor in human neuroblastoma SH-SY5Y cells. J Neurochem,1990,55(4):1390-6
8 Smith EL, Hainsworth AH. Acute effects of interleukin-1 β on noradrenaline release from the human neuroblastoma cell line SH-SY5Y. Neurosci Lett, 1998,251(2):89-92
9 Webster NJ, Vaughan PF, Peers C. Hypoxic enhancement of evoked noradrenaline release from the human neuroblastoma SH-SY5Y. Brain Res Mol Brain Res, 2001,89(1-2):50-7
10 Taglialatela M, Secondo A, Fresi A, et al. Inhibition of depolarization-induced [3H]noradrenaline release from SH-SY5Y human neuroblastoma cells by some second-generation H(1) receptor antagonists through blockade of store-operated Ca2+ channels (SOCs). Biochem Pharmacol, 2001,62(9):1229-38
11 钱淑文,冯琼,朱粹青. β-淀粉样蛋白寡聚体对人神经母细胞瘤SH-SY5Y 细胞 Munc-18 表达的调节.细胞生物学杂志,2005,27(2): 178-182
12 Voigt A, Zintl F. Effects of retinoic acid on proliferation, apoptosis, cytotoxicity, migration, and invasion of neuroblastoma cells. Med Pediatr Oncol, 2003,40(4):205-13
13 Chu PW, Cheung WM, Kwong YL. Differential effects of 9-cis, 13-cis and all-trans retinoic acids on the neuronal differentiation of human neuroblastoma cells. Neuroreport, 2003,14(15):1935-9
14 佟海峡,张继红,陆春伟,等. 全反式维甲酸体外诱导神经母细胞瘤细胞分化的实验研究.中国医学工程, 2007,1(15):37-39
15 Harrison LM, Kastin AJ, Zadina JE. Opiate tolerance and dependence: receptors, G-proteins, and antiopiates. Peptides, 1998,19(9):1603-30
16 Williams JT, Christie MJ, Manzoni O. Cellular and synaptic adaptations mediating opioid dependence. Physiol Rev,2001,81(1):299-343
17 Sharma SK, Nirenberg M, Klee WA. Morphine receptors as regulators of adenylate cyclase activity. Proc Natl Acad Sci USA,1975,72(2):590-4
18 Nestler EJ, Alreja M, Aghajanian GK. Molecular and cellular mechanisms of opiate action: studies in the rat locus coeruleus. Brain Res Bull, 1994,35(5-6):521-8
19 Nestler EJ, Tallman JF. Chronic morphine treatment increases cyclic AMP-dependent protein kinase activity in the rat locus coeruleus. Mol Pharmacol,1988,33(2):127-32
20 Rasmussen K, Beitner-Johnson DB, Krystal JH, et al. Opiate withdrawal and the rat locus coeruleus: behavioral, electrophysiological, and biochemical correlates,1990,10(7):2308-17
21 Terwilliger RZ, Beitner-Johnson D, Sevarino KA, et al. A general role for adaptations in G-proteins and the cyclic AMP system in mediating the chronic actions of morphine and cocaine on neuronal function. Brain Res, 1991,548(1-2):100-10
22 Eriksson PS, Carlsson B, Isaksson OG, et al. Altered amounts of G-protein mRNA and cAMP accumulation after long-term opioid receptor stimulation of neurons in primary culture from the rat cerebral cortex. Brain Res Mol Brain Res, 1992,14(4):317-25
23 Duman RS, Tallman JF, Nestler EJ. Acute and chronic opiate-regulation of adenylate cyclase in brain: specific effects in locus coeruleus. J Pharmacol Exp Ther,1988,246(3):1033-9
24 Nestler EJ. Molecular basis of long-term plasticity underlying addiction. Nat Rev Neurosci,2001,2(2):119-28
25 Traber J, Fischer K, Latzin S, et al. Morphine antagonises action of prostaglandin in neuroblastoma and neuroblastoma times glioma hybrid cells. Nature, 1975,253(5487):120-2
26 Sharma SK, Klee WA, Nirenberg M. Opiate-dependent modulation of adenylate cyclase. Proc Natl Acad Sci U S A,1977,74(8):3365-9
27 Law PY, Koehler JE, Loh HH. Comparison of opiate inhibition of adenylate cyclase activity in neuroblastoma N18tG2 and neuroblastoma x glioma NG108-15 hybrid cell lines. Mol Pharmacol,1982,21(2):483-91
28 Kazmi SM, Mishra RK. Comparative pharmacological properties and functional coupling of mu and delta opioid receptor sites in human neuroblastoma SH-SY5Y cells. Mol Pharmacol,1987,32(1):109-18
29 Ammer H, Schulz R. Morphine dependence in human neuroblastoma SH-SY5Y cells is associated with adaptive changes in both the quantity and functional interaction of PGE1 receptors and stimulatory G proteins. Brain Res,1996,707(2):235-44
30 Wang Z, Bilsky EJ, Porreca F, et al. Constitutive mu opioid receptor activation as a regulatory mechanism underlying narcotic tolerance and dependence. Life Sci, 1994,54(20):PL339-50
31 Wang Z, Sadee W. Tolerance to morphine at the mu-opioid receptor differentially induced by cAMP-dependent protein kinase activation and morphine. Eur J Pharmacol,2000,389(2-3):165-71
32 Frey EA, Kebabian JW. A mu-opiate receptor in 7315c tumor tissue mediates inhibition of immunoreactive prolactin release and adenylate cyclase activity. Endocrinology,1984,115(5):1797-804
33 Puttfarcken PS, Werling LL, Cox BM. Effects of chronic morphine exposure on opioid inhibition of adenylyl cyclase in 7315c cell membranes: a useful model for the study of tolerance at mu opioid receptors. Mol Pharmacol,1988,33(5):520-7
34 Olasmaa M, Terenius L. Prolonged exposure of human neuroblastoma SH-SY5Y cell line to morphine and oxotremorine modulates signal transduction in adenylate cyclase system. Brain Res, 1988,475(2):291-6
1 殷彬,臧德馨. 阿片类药物对神经递质的影响. 国外医学精神病学分册,1997,24(2):85-88
2 van Ree JM, Gerrits MA, Vanderschuren LJ. Opioids, reward and addiction: An encounter of biology, psychology, and medicine. Pharmacol Rev, 1999,51:341-96
3 De Vries TJ, Shippenberg TS. Neural systems underlying opiate addiction. J Neurosci, 2002,22:3321-5
4 张战赛,林兴崇. 阿片类药物的条件性位置偏爱效应及其奖赏机制. 中国心理卫生杂志,2004,18(12):868-871
5 Meririnne E, Kankaanpaa A, Lillsunde P, et al. The effects of diazepam and zolpidem on cocaine- and amphetamine-induced place preference. Pharmacol Biochem Behav, 1999,62:159-64
6 徐美林,余娟,陈崇宏. 奥丹西隆对小鼠吗啡条件性位置偏爱效应的影响. 中国药物依赖性杂志,2004,13(2):105-108
7 Gerra G, Zaimovic A, Moi G, et al. Aggressive responding in abstinent heroin addicts: neuroendocrine and personality correlates. Prog Neuropsychopharmacol Biol Psychiatry, 2004, 28(1):129-39
8 Miao H, Qin By, Yang Y, et al. Ultrastructural changes in rat locus coeruleus induced by chronic opioids. Acta Neuropatho, 1997;94:109
9 Kamei J, Ohsawa M. Role of noradrenergic functions in the modification of naloxone-precipitated withdrawal jumping in morphine-dependent mice by diabetes. Life Sci, 1997, 60(15):PL223 -8
10 Tomkin DM, Sellers EM. Addition and the brain:the role of neurotransmitters in the cause and treatment of drug dependence. CMAJ,2001,164(6):817-821
11 Spanagel R, Almeida OF, Bartl C, et al. Endogenous kappa-opioid systems in opiate withdrawal: role in aversion and accompanying changes in mesolimbic dopamine release. Psychopharmacology (Berl), 1994,115(1-2):121-7
12 Walker JM, Thompson LA, Frascella J, et al. Opposite effects of mu and kappa opiates on the firing-rate of dopamine cells in the substantia nigra of the rat. Eur J Pharmacol, 1987,134(1):53-9
13 Kiyatkin EA, Wise RA, Gratton A. Drug- and behavior-associated changes in dopamine-related electrochemical signals during intravenous heroin self-administration in rats. Synapse, 1993,14(1):60-72
14 Smolders I, De Klippel N, Sarre S, et al. Tonic GABA-ergic modulation of striatal dopamine release studied by in vivo microdialysis in the freely moving rat. Eur J Pharmacol, 1995,284(1-2):83-91
15 Celsen B, Kuschinsky K. Effects of morphine on kinetics of 14C-dopamine in rat striatal slices. Naunyn Schmiedebergs Arch Pharmacol, 1974,284(2):159-65
16 Kuschinsky K, Celsen B, Huppertz A. Studies on dopamine kinetics in rat brain tissue under the influence of morphine. Int J Clin PharmacolBiopharm, 1975,12(1-2):129-33
17 Marien M, Brien J, Jhamandas K. Regional release of [3H] dopamine from rat brain in vitro: effects of opioids on release induced by potassium, nicotine, and L-glutamic acid. Can J Physiol Pharmacol, 1983,61(1):43-60
18 Westfall TC, Grant H, Naes L, et al. The effect of opioid drugs on the release of dopamine and 5-hydroxytryptamine from rat striatum following activation of nicotinic-cholinergic receptors. Eur J Pharmacol, 1983,92(1-2):35-42
19 Widdowson PS, Holman RB. Ethanol-induced increase in endogenous dopamine release may involve endogenous opiates. J Neurochem, 1992,59(1):157-63
20 Piepponen TP, Mikkola JA, Ruotsalainen M, et al. Characterization of the decrease of extracellular striatal dopamine induced by intrastriatal morphine administration. Br J Pharmacol, 1999,127(1):268-74
21 Saiani L, Guidotti A. Opiate receptor-mediated inhibition of catecholamine release in primary cultures of bovine adrenal chromaffin cells. J Neurochem, 1982,39(6):1669-76
22 Wallington CL, Peters DM, Gelb CR, et al. Orphanin FQ/nociceptin and mu-opioid receptor mRNA levels in human SH-SY5Y neuroblastoma cells: effects of activating the cAMP-PKA signal transduction pathway. Brain Res, 2002,944(1-2):73-81
23 De Souza Silva MA, Mattern C, Hacker R, et al. Intranasal administration of the dopaminergic agonists L-DOPA, amphetamine, and cocaine increases dopamine activity in the neostriatum: a microdialysis study in the rat. J Neurochem, 1997,68(1):233-9
24 Murphy NP, Vaughan PF, Ball SG, et al. The cholinergic regulation of intracellular calcium in the human neuroblastoma, SH-SY5Y. J Neurochem, 1991,57(6):2116-23
25 Seward EP, Henderson G. Characterization of two components of the N-like, high-threshold-activated calcium channel current in differentiated SH-SY5Y cells. Pflugers Arch, 1990,417(2):223-30
26 Bean BP. Neurotransmitter inhibition of neuronal calcium currents by changes in channel voltage dependence. Nature, 1989,340(6229): 153-6
27 Hirning LD, Fox AP, McCleskey EW, et al. Dominant role of N-type Ca2+ channels in evoked release of norepinephrine from sympathetic neurons. Science, 1988,239(4835):57-61
28 Kasai H. Voltage- and time-dependent inhibition of neuronal calcium channels by a GTP-binding protein in a mammalian cell line. J Physiol, 1992,448:189-209
29 Shen KZ, Surprenant A. Noradrenaline, somatostatin and opioids inhibit activity of single HVA/N-type calcium channels in excised neuronal membranes. Pflugers Arch, 1991,418(6):614-6
30 Seyedi N, Mackins CJ, Machida T, et al. Histamine H3-receptor-induced attenuation of norepinephrine exocytosis: a decreased protein kinase a activity mediates a reduction in intracellular calcium. J Pharmacol Exp Ther, 2005,312:272-80
31 Seward E, Hammond C, Henderson G. Mu-opioid-receptor-mediated inhibition of the N-type calcium-channel current. Proc Biol Sci, 1991,244:129-35
32 Seward EP, Henderson G. Characterization of two components of the N-like, high-threshold-activated calcium channel current in differentiated SH-SY5Y cells. Pflugers Arch, 1990,417:223-30
33 Murphy NP, Ball SG, Vaughan PF. Potassium- and carbachol-evoked release of [3H]noradrenaline from human neuroblastoma cells, SH-SY5Y. J Neurochem, 1991,56:1810-5
34 Vaughan PF, Kaye DF, Reeve HL, et al. Nicotinic receptor-mediated release of noradrenaline in the human neuroblastoma SH-SY5Y. J Neurochem, 1993,60:2159-66
35 Turner NA, Walker JH, Ball SG, et al. Phorbol ester-enhanced noradrenaline secretion correlates with the presence and activity of protein kinase C-alpha in human SH-SY5Y neuroblastoma cells. J Neurochem, 1996,66:2381-9
36 Levi R, Seyedi N, Schaefer U, et al. Histamine H3-receptor signaling in cardiac sympathetic nerves: Identification of a novel MAPK-PLA2-COX-PGE2-EP3R pathway. Biochem Pharmacol, 2007,73:1146-56
37 Yang Y, Liu X, Ding JH, et al. Effects of iptakalim on rotenone-induced cytotoxicity and dopamine release from PC12 cells. Neurosci Lett, 2004,366(1):53-7
1 DeBello WM, O'Connor V, Dresbach T, et al. SNAP-mediated protein-protein interactions essential for neurotransmitter release. Nature, 1995,373(6515):626-30
2 Sudlow AW, McFerran BW, Bodill H, et al. Similar effects of α- and β-SNAP on Ca2+-regulated exocytosis. FEBS Lett, 1996,393(2-3):185-8
3 郑芳,吴群,姜小丹,等. NK92 细胞的胞吐效应与 SNAP-23 蛋白转位相关. 中华微生物学和免疫学杂志,2005,2(25) :85-88
4 Borowsky B, Hoffman BJ. Neurotransmitter transporters: molecular biology, function, and regulation. Int Rev Neurobiol, 1995,38:139-99
5 Bonisch H, Bruss M. The noradrenaline transporter of the neuronal plasma membrane. Ann N Y Acad Sci, 1994,733:193-202
6 曹君利,丁海雷,杨国栋,等. 囊泡胞吐机制及与其相关的神经元可塑性. 生命科学,2003,15:283-288
7 Brose N, Petrenko AG, Sudhof TC, et al. Synaptotagmin: a calcium sensor on the synaptic vesicle surface. Science, 1992,256:1021-5
8 Adolfsen B, Littleton JT. Genetic and molecular analysis of the synaptotagmin family. Cell Mol Life Sci, 2001,58:393-402
9 Lonart G. RIM1: an edge for presynaptic plasticity. Trends Neurosci, 2002,25:329-32
10 Rizo J, Sudhof TC. Snares and Munc18 in synaptic vesicle fusion. Nat Rev Neurosci, 2002,3:641-53
11 Betz A, Thakur P, Junge HJ, et al. Functional interaction of the active zone proteins Munc13-1 and RIM1 in synaptic vesicle priming. Neuron, 2001,30:183-96
12 Morgan A, Dimaline R, Burgoyne RD. The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. J Biol Chem, 1994,269:29347-50
13 Griff IC, Schekman R, Rothman JE, et al. The yeast SEC17 gene product is functionally equivalent to mammalian α-SNAP protein. J Biol Chem, 1992,267(17):12106-15
14 Peter F, Wong SH, Subramaniam VN, et al. Α-SNAP but not γ-SNAP is required for ER-Golgi transport after vesicle budding and the Rab1-requiring step but before the EGTA-sensitive step. J Cell Sci, 1998,111 ( Pt 17):2625-33
15 Reuveny E, Narahashi T. Two types of high voltage-activated calcium channels in SH-SY5Y human neuroblastoma cells. Brain Res, 1993,603:64-73
16 Purkiss JR, Friis LM, Doward S, et al. Clostridium botulinum neurotoxinsact with a wide range of potencies on SH-SY5Y human neuroblastoma cells. Neurotoxicology, 2001,22:447-53.
17 Paddock SW. Principles and practices of laser scanning confocal microscopy. Mol Biotechnol ,2000,16(2): 127-149
18 Kreft M, Stenovec M, Zorec R. Focus-drift correction in time-lapse confocal imaging. Ann NY Acad Sci ,2005,1048(6):321-330
19 Collazo A, Bricaud O, Desai K. Use of confocal microscopy in comparative studies of vertebrate morphology. Methods Enzymol, 2005,395:521-543
20 Seyedi N, Mackins CJ, Machida T, et al. Histamine H3-receptor-induced attenuation of norepinephrine exocytosis: a decreased protein kinase a activity mediates a reduction in intracellular calcium. J Pharmacol Exp Ther, 2005,312:272-80
21 Wang Z, Sadee W. Tolerance to morphine at the mu-opioid receptor differentially induced by cAMP-dependent protein kinase activation and morphine. Eur J Pharmacol,2000,389(2-3):165-71
22 Nestler EJ, Alreja M, Aghajanian GK. Molecular and cellular mechanisms of opiate action: studies in the rat locus coeruleus. Brain Res Bull, 1994,35(5-6):521-8
23 Nestler EJ, Tallman JF. Chronic morphine treatment increases cyclic AMP-dependent protein kinase activity in the rat locus coeruleus. Mol Pharmacol,1988,33(2):127-32
24 Eriksson PS, Carlsson B, Isaksson OG, et al. Altered amounts of G-protein mRNA and cAMP accumulation after long-term opioid receptor stimulation of neurons in primary culture from the rat cerebral cortex. Brain Res Mol Brain Res, 1992,14(4):317-25
25 Duman RS, Tallman JF, Nestler EJ. Acute and chronic opiate-regulation of adenylate cyclase in brain: specific effects in locus coeruleus. J Pharmacol Exp Ther,1988,246(3):1033-9
26 Nestler EJ. Molecular basis of long-term plasticity underlying addiction. Nat Rev Neurosci,2001,2(2):119-28
27 Llinas R, Sugimori M, Silver RB. Microdomains of high calcium concentration in a presynaptic terminal. Science, 1992,256:677-9.
28 Zucker RS. Calcium and transmitter release. J Physiol Paris, 1993,87:25-36
29 Bai J, Wang CT, Richards DA, et al. Fusion pore dynamics are regulated by synaptotagmin*t-SNARE interactions. Neuron, 2004,41:929-42
30 Stenbeck G. Soluble NSF-attachment proteins. Int J Biochem Cell Biol, 1998,30:573-7
31 Nishiki T, Nihonmatsu I, Tsuhara Y, et al. Distribution of soluble N-ethylmaleimide fusion protein attachment proteins (SNAPs) in the rat nervous system. Neuroscience, 2001,107:363-71
32 Sollner T. SNAREs and targeted membrane fusion. FEBS Lett, 1995,369:80-3
33 Woodman PG. The roles of NSF, SNAPs and SNAREs during membrane fusion. Biochim Biophys Acta, 1997,1357:155-72
34 Sheng ZH, Yokoyama CT, Catterall WA. Interaction of the synprint site of N-type Ca2+ channels with the C2B domain of synaptotagmin I. Proc Natl Acad Sci U S A, 1997,94:5405-10
35 Rettig J, Sheng ZH, Kim DK, et al. Isoform-specific interaction of the alpha1A subunits of brain Ca2+ channels with the presynaptic proteins syntaxin and SNAP-25. Proc Natl Acad Sci U S A, 1996,93:7363-8
36 Sheng ZH, Rettig J, Cook T, et al. Calcium-dependent interaction of N-type calcium channels with the synaptic core complex. Nature, 1996,379:451-4
37 Mochida S, Sheng ZH, Baker C, et al. Inhibition of neurotransmission by peptides containing the synaptic protein interaction site of N-type Ca2+ channels. Neuron, 1996,17:781-8
38 杜宝国,张瑞玲,李昌琪,等.吗啡依赖大鼠伏隔核区单胺转运体表达的时点变化.中国药物依赖性杂志,2006,15:358-362
39 Xu NJ, Bao L, Fan HP, et al. Morphine withdrawal increases glutamate uptake and surface expression of glutamate transporter GLT1 at hippocampal synapses. J Neurosci, 2003,23:4775-84
40 Gandhi SP, Stevens CF. Three modes of synaptic vesicular recycling revealed by single-vesicle imaging. Nature, 2003,423:607-13
1 Nishiki T, Nihonmatsu I, Tsuhara Y, et al. Distribution of soluble N-ethylmaleimide fusion protein attachment proteins (SNAPs) in the rat nervous system. Neuroscience, 2001,107:363-71
2 Whiteheart SW, Griff IC, Brunner M, et al. SNAP family of NSF attachment proteins includes a brain-specific isoform. Nature, 1993,362:353-5
3 Stenbeck G. Soluble NSF-attachment proteins. Int J Biochem Cell Biol, 1998,30:573-7
4 Kato K. A Collection of cDNA Clones with Specific Expression Patterns in Mouse Brain. Eur J Neurosci, 1990,2:704-711
5 Rothman JE, Warren G. Implications of the SNARE hypothesis for intracellular membrane topology and dynamics. Curr Biol, 1994,4:220-33
6 Cho SJ, Kelly M, Rognlien KT, et al. SNAREs in opposing bilayers interact in a circular array to form conducting pores. Biophys J, 2002,83:2522-7
7 Cho WJ, Jeremic A, Jena BP. Size of supramolecular SNARE complex: membrane-directed self-assembly. J Am Chem Soc, 2005,127:10156-7
8 Sutton RB, Fasshauer D, Jahn R, et al. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature, 1998,395:347-53
9 Hayashi T, McMahon H, Yamasaki S, et al. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly. EMBO J, 1994,13:5051-61
10 Yang B, Gonzalez L Jr, Prekeris R, et al. SNARE interactions are not selective. Implications for membrane fusion specificity. J Biol Chem, 1999,274:5649-53
11 Hanson PI, Heuser JE, Jahn R. Neurotransmitter release - four years of SNARE complexes. Curr Opin Neurobiol, 1997,7:310-5
12 Brunger AT. Structure and function of SNARE and SNARE-interactingproteins. Q Rev Biophys, 2005,38:1-47
13 Jahn R, Scheller RH. SNAREs--engines for membrane fusion. Nat Rev Mol Cell Biol, 2006,7:631-43
14 Sudhof TC. The synaptic vesicle cycle. Annu Rev Neurosci, 2004,27:509-47
15 Fasshauer D, Margittai M. A transient N-terminal interaction of SNAP-25 and syntaxin nucleates SNARE assembly. J Biol Chem, 2004,279:7613-21
16 Fasshauer D, Otto H, Eliason WK, et al. Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation. J Biol Chem, 1997,272:28036-41
17 Sorensen JB, Wiederhold K, Muller EM, et al. Sequential N- to C-terminal SNARE complex assembly drives priming and fusion of secretory vesicles. EMBO J, 2006,25:955-66
18 Weber T, Zemelman BV, McNew JA, et al. SNAREpins: minimal machinery for membrane fusion. Cell, 1998,92:759-72
19 McNew JA, Weber T, Parlati F, et al. Close is not enough: SNARE-dependent membrane fusion requires an active mechanism that transduces force to membrane anchors. J Cell Biol, 2000,150:105-17
20 Hu C, Ahmed M, Melia TJ, et al. Fusion of cells by flipped SNAREs. Science, 2003,300:1745-9
21 Fix M, Melia TJ, Jaiswal JK, et al. Imaging single membrane fusion events mediated by SNARE proteins. Proc Natl Acad Sci U S A, 2004,101:7311-6
22 Sollner T, Bennett MK, Whiteheart SW, et al. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell, 1993,75:409-18
23 Clary DO, Griff IC, Rothman JE. SNAPs, a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast. Cell, 1990,61:709-21
24 Wilson DW, Whiteheart SW, Wiedmann M, et al. A multisubunit particle implicated in membrane fusion. J Cell Biol, 1992,117:531-8
25 Morgan A, Dimaline R, Burgoyne RD. The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. J Biol Chem, 1994,269:29347-50
26 Matveeva E, Whiteheart SW. The effects of SNAP/SNARE complexes on the ATPase of NSF. FEBS Lett, 1998,435:211-4
27 Hayashi T, Yamasaki S, Nauenburg S, et al. Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. EMBO J, 1995,14:2317-25
28 McMahon HT, Sudhof TC. Synaptic core complex of synaptobrevin, syntaxin, and SNAP-25 forms high affinity alpha-SNAP binding site. J Biol Chem, 1995,270:2213-7
29 Tagaya M, Wilson DW, Brunner M, et al. Domain structure of an N-ethylmaleimide-sensitive fusion protein involved in vesicular transport. J Biol Chem, 1993,268:2662-6
30 Whiteheart SW, Brunner M, Wilson DW, et al. Soluble N-ethylmaleimide-sensitive fusion attachment proteins (SNAPs) bind to a multi-SNAP receptor complex in Golgi membranes. J Biol Chem, 1992,267:12239-43
31 Wimmer C, Hohl TM, Hughes CA, et al. Molecular mass, stoichiometry, and assembly of 20 S particles. J Biol Chem, 2001,276:29091-7
32 Hanson PI, Otto H, Barton N, et al. The N-ethylmaleimide-sensitive fusion protein and alpha-SNAP induce a conformational change in syntaxin. J Biol Chem, 1995,270:16955-61
33 Barnard RJ, Morgan A, Burgoyne RD. Domains of alpha-SNAP required for the stimulation of exocytosis and for N-ethylmalemide-sensitive fusion protein (NSF) binding and activation. Mol Biol Cell, 1996,7:693-701
34 Barnard RJ, Morgan A, Burgoyne RD. Stimulation of NSF ATPase activity by alpha-SNAP is required for SNARE complex disassembly andexocytosis. J Cell Biol, 1997,139:875-83
35 Tani K, Shibata M, Kawase K, et al. Mapping of functional domains of gamma-SNAP. J Biol Chem, 2003,278:13531-8
36 Puschel AW, O'Connor V, Betz H. The N-ethylmaleimide-sensitive fusion protein (NSF) is preferentially expressed in the nervous system. FEBS Lett, 1994,347:55-8
37 DeBello WM, O'Connor V, Dresbach T, et al. SNAP-mediated protein-protein interactions essential for neurotransmitter release. Nature JT - Nature, 1995,373:626-30
38 Hirling H, Scheller RH. Phosphorylation of synaptic vesicle proteins: modulation of the alpha SNAP interaction with the core complex. Proc Natl Acad Sci U S A, 1996,93:11945-9
39 DiAntonio A, Parfitt KD, Schwarz TL. Synaptic transmission persists in synaptotagmin mutants of Drosophila. Cell, 1993,73:1281-90
40 Littleton JT, Stern M, Schulze K, et al. Mutational analysis of Drosophila synaptotagmin demonstrates its essential role in Ca(2+)-activated neurotransmitter release. Cell, 1993,74:1125-34
41 DiAntonio A, Schwarz TL. The effect on synaptic physiology of synaptotagmin mutations in Drosophila. Neuron, 1994,12:909-20
42 Broadie K, Bellen HJ, DiAntonio A, et al. Absence of synaptotagmin disrupts excitation-secretion coupling during synaptic transmission. Proc Natl Acad Sci U S A, 1994,91:10727-31
43 Griff IC, Schekman R, Rothman JE, et al. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein. J Biol Chem JT, 1992,267:12106-15
44 Schiavo G, Gmachl MJ, Stenbeck G, et al. A possible docking and fusion particle for synaptic transmission. Nature, 1995,378:733-6
45 Martin HG, Henley JM, Meyer G. Novel putative targets of N-ethylmaleimide sensitive fusion protein (NSF) and alpha/beta soluble NSF attachment proteins (SNAPs) include the Pak-binding nucleotide exchange factor betaPIX. J Cell Biochem, 2006,99:1203 -15
46 Peter F, Wong SH, Subramaniam VN, et al. Alpha-SNAP but not gamma-SNAP is required for ER-Golgi transport after vesicle budding and the Rab1-requiring step but before the EGTA-sensitive step. J Cell Sci , 1998,111 ( Pt 17):2625-33
47 Nagamatsu S, Watanabe T, Nakamichi Y, et al. alpha-soluble N-ethylmaleimide-sensitive factor attachment protein is expressed in pancreatic beta cells and functions in insulin but not gamma-aminobutyric acid secretion. J Biol Chem,1999,274:8053 -60
48 Morgan A, Burgoyne RD. A role for soluble NSF attachment proteins (SNAPs) in regulated exocytosis in adrenal chromaffin cells. EMBO J, 1995,14:232-9
49 Abonyo BO, Wang P, Narasaraju TA, et al. Characterization of alpha-soluble N-ethylmaleimide-sensitive fusion attachment protein in alveolar type II cells: implications in lung surfactant secretion. Am J Respir Cell Mol Biol, 2003,29:273-82
50 Baur T, Ramadan K, Schlundt A, et al. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts. J Cell Sci, 2007,120:2895-903
51 Echard A, Hickson GR, Foley E, et al. Terminal cytokinesis events uncovered after an RNAi screen. Curr Biol, 2004,14:1685-93
52 Sudlow AW, McFerran BW, Bodill H, et al. Similar effects of alpha- and beta-SNAP on Ca2+-regulated exocytosis. FEBS Lett, 1996,393:185-8
53 Xu J, Xu Y, Ellis-Davies GC, et al. Differential regulation of exocytosis by alpha- and beta-SNAPs. J Neurosci, 2002,22:53-61
54 Andreeva AV, Kutuzov MA, Voyno-Yasenetskaya TA. A ubiquitous membrane fusion protein alpha SNAP: a potential therapeutic target for cancer, diabetes and neurological disorders? Expert Opin Ther Targets, 2006,10:723-33
55 Yoo BC, Cairns N, Fountoulakis M, et al. Synaptosomal proteins, beta-soluble N-ethylmaleimide-sensitive factor attachment protein(beta-SNAP), gamma-SNAP and synaptotagmin I in brain of patients with Down syndrome and Alzheimer's disease. Dement Geriatr Cogn Disord, 2001,12:219-25
56 Sultana R, Boyd-Kimball D, Poon HF, et al. Redox proteomics identification of oxidized proteins in Alzheimer's disease hippocampus and cerebellum: an approach to understand pathological and biochemical alterations in AD. Neurobiol Aging, 2006,27:1564-76
57 Weller AE, Dahl JP, Lohoff FW, et al. Analysis of variations in the NAPG gene on chromosome 18p11 in bipolar disorder. Psychiatr Genet, 2006,16:3-8
2 Nestler EJ, Aghajanian GK. Molecular and cellular basis of addiction. Science (New York, N.Y.), 1997, 278(5335):58-63
3 Williams JT, Christie MJ, Manzoni O. Cellular and synaptic adaptations mediating opioid dependence. Physiol Rev, 2001, 81(1):299-343
4 Taylor DA, Fleming WW. Unifying perspectives of the mechanisms underlying the development of tolerance and physical dependence to opioids. J Pharmacol Exp Ther, 2001, 297(1):11-18
5 曹君利,丁海雷,杨国栋,等.囊泡胞吐机制及与其相关的神经元可塑性.生命科学,2003,5(15): 283-288
6 蔡倩,陆佩华,盛祖抗.神经末梢突触囊泡神经递质过程的调控蛋白.生理科学进展,2003,1(34): 6-10
7 王堰婷,陆佩华,盛祖抗.神经递质释放过程中的可溶性 NSF 附着蛋白受体(SNARE)和相关蛋白.生物化学与生物物理进展,2002,29(1): 35-38
8 Sollner T, Whiteheart SW, Brunner M, et al. SNAP receptors implicated in vesicle targeting and fusion. Nature, 1993, 362(6418):318-24
9 Clary DO, Rothman JE. Purification of three related peripheral membrane proteins needed for vesicular transport. J Biol Chem, 1990,265(17): 10109-17
10 Whiteheart SW, Griff IC, Brunner M, et al. SNAP family of NSFattachment proteins includes a brain-specific isoform. Nature, 1993, 362(6418):353-5
11 Haynes LP, Barnard RJ, Morgan A, et al. Stimulation of NSF ATPase activity during t-SNARE priming. FEBS Lett, 1998, 436(1):1-5
12 Morgan A, Dimaline R, Burgoyne RD. The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. J Biol Chem, 1994, 269(47):29347-50
13 De Souza Silva MA, Mattern C, Hacker R, et al. Intranasal administration of the dopaminergic agonists L-DOPA, amphetamine, and cocaine increases dopamine activity in the neostriatum: a microdialysis study in the rat. J Neurochem, 1997, 68(1):233-9
14 Suzuki T, Tsuji M, Mori T, et al. Effect of naltrindole on the development of physical dependence on morphine in mice: a behavioral and biochemical study. Life Sci, 1995, 57(17):PL247-52
15 Wise RA, Bozarth MA. A psychomotor stimulant theory of addiction. Psychol Rev, 1987, 94(4):469-92
16 Leone P, Pocock D, Wise RA. Morphine-dopamine interaction: ventral tegmental morphine increases nucleus accumbens dopamine release. Pharmacol Biochem Behav, 1991, 39(2):469 -72
17 Di Chiara G, Imperato A. Drugs abused by humans preferentially increase synaptic dopamine concentrations in the mesolimbic system of freely moving rats. Proc Natl Acad Sci U S A, 1988, 85(14):5274-8
18 Gerra G, Zaimovic A, Moi G, et al. Aggressive responding in abstinent heroin addicts: neuroendocrine and personality correlates. Prog Neuropsychopharmacol Biol Psychiatry, 2004, 28(1):129-39
19 Miao H, Qin By, Yang Y, et al. Ultrastructural changes in rat locus coeruleus induced by chronic opioids. Acta Neuropatho, 1997;94:109
20 Kamei J, Ohsawa M. Role of noradrenergic functions in the modification of naloxone-precipitated withdrawal jumping in morphine-dependent mice by diabetes. Life Sci, 1997, 60(15):PL223 -8
21 Mylecharane EJ. Ventral tegmental area 5-HT receptors: mesolimbicdopamine release and behavioural studies. Behav Brain Res, 1996, 73 (1-2):1-5
22 方芳,曹清,宋福津,等.吗啡长时程对 SH-SY5Y 细胞 cAMP 系统及c-Fos 磷酸化的影响.中国医学科学学报,1999,4(21): 262-267
23 Xu NJ, Yu YX, Zhu JM, et al. Inhibition of SNAP - 25 phosphorylation at Ser187 is involved in chronic morphine - induced down - regulation of SNARE complex formation [J]. J Biol Chem,2004,279 (39):40601– 40608
24 Manzoni OJ, Williams JT. Presynaptic regulation of glutamate release in the ventral tegmental area during morphine withdrawal [J]. J Neurosci, 1999, 19:6629–6636
25 Sepulveda MJ, Hernandez L, Rada P, et al. Effect of precipitated withdrawal on extracellular glutamate and aspartate in the nucleus accumbens of chronically morphine-treated rats: an in vivo microdialysis study [J]. Pharmacol Biochem Behav, 1998, 60:255–262
26 Sudlow AW, McFerran BW, Bodill H, Barnard RJ, Morgan A, Burgoyne RD.Similar effects of α- and β-SNAP on Ca2+-regulated exocytosis. FEBS Lett. 1996 Sep 16;393(2-3):185~8
27 Nagamatsu S, Watanabe T, Nakamichi Y, et al. α-soluble N-ethylmaleimide-sensitive factor attachment protein is expressed in pancreatic β cells and functions in insulin but not γ-aminobutyric acid secretion. J Biol Chem, 1999, 274(12):8053-60
28 Abonyo BO, Wang P, Narasaraju TA, et al. Characterization of α-Soluble N-Ethylmaleimide–Sensitive Fusion Attachment Protein in Alveolar Type II Cells: Implications in Lung Surfactant Secretion. Am J Respir Cell Mol Biol, 2003,29:273–282
29 Baur T, Ramadan K, Schlundt A, et al. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts. J Cell Sci, 2007, 120:2895-903
30 George Paxinos,Charles Watson,主编. 诸葛启钏,主译.大鼠脑立体定位图谱[M]. 第 3 版,北京:人民卫生出版社,2005
31 DeBello WM, O'Connor V, Dresbach T, et al. SNAP-mediated protein-protein interactions essential for neurotransmitter release. Nature, 1995, 373(6515):626-30
32 Morgan A, Burgoyne RD. A role for soluble NSF attachment proteins (SNAPs) in regulated exocytosis in adrenal chromaffin cells. EMBO, 1995, 14(2):232-9
33 Xu J, Xu Y, Ellis-Davies GC,et al. Differential Regulation of Exocytosis by α- and β-SNAPs [J]. J of Neurosci,2002,22(1):53-61
34 Nakajima K, Hirose H, Taniguchi M, et al. Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion. EMBO J, 2004, 23:3216-26
35 Schiavo G, Gmachl MJ, Stenbeck G, et al. A possible docking and fusion particle for synaptic transmission. Nature, 1995, 378:733-6
36 Hanley JG, Khatri L, Hanson PI, et al. NSF ATPase and α-/β-SNAPs disassemble the AMPA receptor-PICK1 complex. Neuron, 2002, 34:53-67
37 Chen D, Xu W, He P, et al. Gaf-1, a γ -SNAP-binding protein associated with the mitochondria. J Biol Chem, 2001, 276:13127-35
38 Tani K, Shibata M, Kawase K, et al. Mapping of functional domains of γ-SNAP. J Biol Chem, 2003, 278:13531-8
39 Nakamichi Y, Nagamatsu S. α-SNAP functions in insulin exocytosis from mature, but not immature secretory granules in pancreatic β cells [J]. Biochem Biophys Res Commun,1999,260(1):127-132
1 Yu VC, Richards ML, Sadee W. A human neuroblastoma cell line expresses mu and delta opioid receptor sites. J Biol Chem, 1986,261(3):1065-70
2 Yu VC, Sadee W. Efficacy and tolerance of narcotic analgesics at the mu opioid receptor in differentiated human neuroblastoma cells. J Pharmacol Exp Ther, 1988,245(1):350-5
3 Odelstad L, Pahlman S, Nilsson K, et al. Neuron-specific enolase in relation to differentiation in human neuroblastoma. Brain Res, 1981,224(1):69-82
4 Zadina JE, Chang SL, Ge LJ, et al. Mu opiate receptor down-regulation by morphine and up-regulation by naloxone in SH-SY5Y human neuroblastoma cells. J Pharmacol Exp Ther, 1993,265(1):254-62
5 Kam AY, Chan AS, Wong YH. Phosphatidylinositol-3 kinase is distinctively required for mu-, but not kappa-opioid receptor-induced activation of c-Jun N-terminal kinase. J Neurochem,2004,89(2):391-402
6 Hou YN, Vlaskovska M, Cebers G, et al. A mu-receptor opioid agonist induces AP-1 and NF-kappa B transcription factor activity in primary cultures of rat cortical neurons. Neurosci Lett, 1996,212(3):159-62
7 Yu VC, Eiger S, Duan DS, et al. Regulation of cyclic AMP by the mu-opioid receptor in human neuroblastoma SH-SY5Y cells. J Neurochem,1990,55(4):1390-6
8 Smith EL, Hainsworth AH. Acute effects of interleukin-1 β on noradrenaline release from the human neuroblastoma cell line SH-SY5Y. Neurosci Lett, 1998,251(2):89-92
9 Webster NJ, Vaughan PF, Peers C. Hypoxic enhancement of evoked noradrenaline release from the human neuroblastoma SH-SY5Y. Brain Res Mol Brain Res, 2001,89(1-2):50-7
10 Taglialatela M, Secondo A, Fresi A, et al. Inhibition of depolarization-induced [3H]noradrenaline release from SH-SY5Y human neuroblastoma cells by some second-generation H(1) receptor antagonists through blockade of store-operated Ca2+ channels (SOCs). Biochem Pharmacol, 2001,62(9):1229-38
11 钱淑文,冯琼,朱粹青. β-淀粉样蛋白寡聚体对人神经母细胞瘤SH-SY5Y 细胞 Munc-18 表达的调节.细胞生物学杂志,2005,27(2): 178-182
12 Voigt A, Zintl F. Effects of retinoic acid on proliferation, apoptosis, cytotoxicity, migration, and invasion of neuroblastoma cells. Med Pediatr Oncol, 2003,40(4):205-13
13 Chu PW, Cheung WM, Kwong YL. Differential effects of 9-cis, 13-cis and all-trans retinoic acids on the neuronal differentiation of human neuroblastoma cells. Neuroreport, 2003,14(15):1935-9
14 佟海峡,张继红,陆春伟,等. 全反式维甲酸体外诱导神经母细胞瘤细胞分化的实验研究.中国医学工程, 2007,1(15):37-39
15 Harrison LM, Kastin AJ, Zadina JE. Opiate tolerance and dependence: receptors, G-proteins, and antiopiates. Peptides, 1998,19(9):1603-30
16 Williams JT, Christie MJ, Manzoni O. Cellular and synaptic adaptations mediating opioid dependence. Physiol Rev,2001,81(1):299-343
17 Sharma SK, Nirenberg M, Klee WA. Morphine receptors as regulators of adenylate cyclase activity. Proc Natl Acad Sci USA,1975,72(2):590-4
18 Nestler EJ, Alreja M, Aghajanian GK. Molecular and cellular mechanisms of opiate action: studies in the rat locus coeruleus. Brain Res Bull, 1994,35(5-6):521-8
19 Nestler EJ, Tallman JF. Chronic morphine treatment increases cyclic AMP-dependent protein kinase activity in the rat locus coeruleus. Mol Pharmacol,1988,33(2):127-32
20 Rasmussen K, Beitner-Johnson DB, Krystal JH, et al. Opiate withdrawal and the rat locus coeruleus: behavioral, electrophysiological, and biochemical correlates,1990,10(7):2308-17
21 Terwilliger RZ, Beitner-Johnson D, Sevarino KA, et al. A general role for adaptations in G-proteins and the cyclic AMP system in mediating the chronic actions of morphine and cocaine on neuronal function. Brain Res, 1991,548(1-2):100-10
22 Eriksson PS, Carlsson B, Isaksson OG, et al. Altered amounts of G-protein mRNA and cAMP accumulation after long-term opioid receptor stimulation of neurons in primary culture from the rat cerebral cortex. Brain Res Mol Brain Res, 1992,14(4):317-25
23 Duman RS, Tallman JF, Nestler EJ. Acute and chronic opiate-regulation of adenylate cyclase in brain: specific effects in locus coeruleus. J Pharmacol Exp Ther,1988,246(3):1033-9
24 Nestler EJ. Molecular basis of long-term plasticity underlying addiction. Nat Rev Neurosci,2001,2(2):119-28
25 Traber J, Fischer K, Latzin S, et al. Morphine antagonises action of prostaglandin in neuroblastoma and neuroblastoma times glioma hybrid cells. Nature, 1975,253(5487):120-2
26 Sharma SK, Klee WA, Nirenberg M. Opiate-dependent modulation of adenylate cyclase. Proc Natl Acad Sci U S A,1977,74(8):3365-9
27 Law PY, Koehler JE, Loh HH. Comparison of opiate inhibition of adenylate cyclase activity in neuroblastoma N18tG2 and neuroblastoma x glioma NG108-15 hybrid cell lines. Mol Pharmacol,1982,21(2):483-91
28 Kazmi SM, Mishra RK. Comparative pharmacological properties and functional coupling of mu and delta opioid receptor sites in human neuroblastoma SH-SY5Y cells. Mol Pharmacol,1987,32(1):109-18
29 Ammer H, Schulz R. Morphine dependence in human neuroblastoma SH-SY5Y cells is associated with adaptive changes in both the quantity and functional interaction of PGE1 receptors and stimulatory G proteins. Brain Res,1996,707(2):235-44
30 Wang Z, Bilsky EJ, Porreca F, et al. Constitutive mu opioid receptor activation as a regulatory mechanism underlying narcotic tolerance and dependence. Life Sci, 1994,54(20):PL339-50
31 Wang Z, Sadee W. Tolerance to morphine at the mu-opioid receptor differentially induced by cAMP-dependent protein kinase activation and morphine. Eur J Pharmacol,2000,389(2-3):165-71
32 Frey EA, Kebabian JW. A mu-opiate receptor in 7315c tumor tissue mediates inhibition of immunoreactive prolactin release and adenylate cyclase activity. Endocrinology,1984,115(5):1797-804
33 Puttfarcken PS, Werling LL, Cox BM. Effects of chronic morphine exposure on opioid inhibition of adenylyl cyclase in 7315c cell membranes: a useful model for the study of tolerance at mu opioid receptors. Mol Pharmacol,1988,33(5):520-7
34 Olasmaa M, Terenius L. Prolonged exposure of human neuroblastoma SH-SY5Y cell line to morphine and oxotremorine modulates signal transduction in adenylate cyclase system. Brain Res, 1988,475(2):291-6
1 殷彬,臧德馨. 阿片类药物对神经递质的影响. 国外医学精神病学分册,1997,24(2):85-88
2 van Ree JM, Gerrits MA, Vanderschuren LJ. Opioids, reward and addiction: An encounter of biology, psychology, and medicine. Pharmacol Rev, 1999,51:341-96
3 De Vries TJ, Shippenberg TS. Neural systems underlying opiate addiction. J Neurosci, 2002,22:3321-5
4 张战赛,林兴崇. 阿片类药物的条件性位置偏爱效应及其奖赏机制. 中国心理卫生杂志,2004,18(12):868-871
5 Meririnne E, Kankaanpaa A, Lillsunde P, et al. The effects of diazepam and zolpidem on cocaine- and amphetamine-induced place preference. Pharmacol Biochem Behav, 1999,62:159-64
6 徐美林,余娟,陈崇宏. 奥丹西隆对小鼠吗啡条件性位置偏爱效应的影响. 中国药物依赖性杂志,2004,13(2):105-108
7 Gerra G, Zaimovic A, Moi G, et al. Aggressive responding in abstinent heroin addicts: neuroendocrine and personality correlates. Prog Neuropsychopharmacol Biol Psychiatry, 2004, 28(1):129-39
8 Miao H, Qin By, Yang Y, et al. Ultrastructural changes in rat locus coeruleus induced by chronic opioids. Acta Neuropatho, 1997;94:109
9 Kamei J, Ohsawa M. Role of noradrenergic functions in the modification of naloxone-precipitated withdrawal jumping in morphine-dependent mice by diabetes. Life Sci, 1997, 60(15):PL223 -8
10 Tomkin DM, Sellers EM. Addition and the brain:the role of neurotransmitters in the cause and treatment of drug dependence. CMAJ,2001,164(6):817-821
11 Spanagel R, Almeida OF, Bartl C, et al. Endogenous kappa-opioid systems in opiate withdrawal: role in aversion and accompanying changes in mesolimbic dopamine release. Psychopharmacology (Berl), 1994,115(1-2):121-7
12 Walker JM, Thompson LA, Frascella J, et al. Opposite effects of mu and kappa opiates on the firing-rate of dopamine cells in the substantia nigra of the rat. Eur J Pharmacol, 1987,134(1):53-9
13 Kiyatkin EA, Wise RA, Gratton A. Drug- and behavior-associated changes in dopamine-related electrochemical signals during intravenous heroin self-administration in rats. Synapse, 1993,14(1):60-72
14 Smolders I, De Klippel N, Sarre S, et al. Tonic GABA-ergic modulation of striatal dopamine release studied by in vivo microdialysis in the freely moving rat. Eur J Pharmacol, 1995,284(1-2):83-91
15 Celsen B, Kuschinsky K. Effects of morphine on kinetics of 14C-dopamine in rat striatal slices. Naunyn Schmiedebergs Arch Pharmacol, 1974,284(2):159-65
16 Kuschinsky K, Celsen B, Huppertz A. Studies on dopamine kinetics in rat brain tissue under the influence of morphine. Int J Clin PharmacolBiopharm, 1975,12(1-2):129-33
17 Marien M, Brien J, Jhamandas K. Regional release of [3H] dopamine from rat brain in vitro: effects of opioids on release induced by potassium, nicotine, and L-glutamic acid. Can J Physiol Pharmacol, 1983,61(1):43-60
18 Westfall TC, Grant H, Naes L, et al. The effect of opioid drugs on the release of dopamine and 5-hydroxytryptamine from rat striatum following activation of nicotinic-cholinergic receptors. Eur J Pharmacol, 1983,92(1-2):35-42
19 Widdowson PS, Holman RB. Ethanol-induced increase in endogenous dopamine release may involve endogenous opiates. J Neurochem, 1992,59(1):157-63
20 Piepponen TP, Mikkola JA, Ruotsalainen M, et al. Characterization of the decrease of extracellular striatal dopamine induced by intrastriatal morphine administration. Br J Pharmacol, 1999,127(1):268-74
21 Saiani L, Guidotti A. Opiate receptor-mediated inhibition of catecholamine release in primary cultures of bovine adrenal chromaffin cells. J Neurochem, 1982,39(6):1669-76
22 Wallington CL, Peters DM, Gelb CR, et al. Orphanin FQ/nociceptin and mu-opioid receptor mRNA levels in human SH-SY5Y neuroblastoma cells: effects of activating the cAMP-PKA signal transduction pathway. Brain Res, 2002,944(1-2):73-81
23 De Souza Silva MA, Mattern C, Hacker R, et al. Intranasal administration of the dopaminergic agonists L-DOPA, amphetamine, and cocaine increases dopamine activity in the neostriatum: a microdialysis study in the rat. J Neurochem, 1997,68(1):233-9
24 Murphy NP, Vaughan PF, Ball SG, et al. The cholinergic regulation of intracellular calcium in the human neuroblastoma, SH-SY5Y. J Neurochem, 1991,57(6):2116-23
25 Seward EP, Henderson G. Characterization of two components of the N-like, high-threshold-activated calcium channel current in differentiated SH-SY5Y cells. Pflugers Arch, 1990,417(2):223-30
26 Bean BP. Neurotransmitter inhibition of neuronal calcium currents by changes in channel voltage dependence. Nature, 1989,340(6229): 153-6
27 Hirning LD, Fox AP, McCleskey EW, et al. Dominant role of N-type Ca2+ channels in evoked release of norepinephrine from sympathetic neurons. Science, 1988,239(4835):57-61
28 Kasai H. Voltage- and time-dependent inhibition of neuronal calcium channels by a GTP-binding protein in a mammalian cell line. J Physiol, 1992,448:189-209
29 Shen KZ, Surprenant A. Noradrenaline, somatostatin and opioids inhibit activity of single HVA/N-type calcium channels in excised neuronal membranes. Pflugers Arch, 1991,418(6):614-6
30 Seyedi N, Mackins CJ, Machida T, et al. Histamine H3-receptor-induced attenuation of norepinephrine exocytosis: a decreased protein kinase a activity mediates a reduction in intracellular calcium. J Pharmacol Exp Ther, 2005,312:272-80
31 Seward E, Hammond C, Henderson G. Mu-opioid-receptor-mediated inhibition of the N-type calcium-channel current. Proc Biol Sci, 1991,244:129-35
32 Seward EP, Henderson G. Characterization of two components of the N-like, high-threshold-activated calcium channel current in differentiated SH-SY5Y cells. Pflugers Arch, 1990,417:223-30
33 Murphy NP, Ball SG, Vaughan PF. Potassium- and carbachol-evoked release of [3H]noradrenaline from human neuroblastoma cells, SH-SY5Y. J Neurochem, 1991,56:1810-5
34 Vaughan PF, Kaye DF, Reeve HL, et al. Nicotinic receptor-mediated release of noradrenaline in the human neuroblastoma SH-SY5Y. J Neurochem, 1993,60:2159-66
35 Turner NA, Walker JH, Ball SG, et al. Phorbol ester-enhanced noradrenaline secretion correlates with the presence and activity of protein kinase C-alpha in human SH-SY5Y neuroblastoma cells. J Neurochem, 1996,66:2381-9
36 Levi R, Seyedi N, Schaefer U, et al. Histamine H3-receptor signaling in cardiac sympathetic nerves: Identification of a novel MAPK-PLA2-COX-PGE2-EP3R pathway. Biochem Pharmacol, 2007,73:1146-56
37 Yang Y, Liu X, Ding JH, et al. Effects of iptakalim on rotenone-induced cytotoxicity and dopamine release from PC12 cells. Neurosci Lett, 2004,366(1):53-7
1 DeBello WM, O'Connor V, Dresbach T, et al. SNAP-mediated protein-protein interactions essential for neurotransmitter release. Nature, 1995,373(6515):626-30
2 Sudlow AW, McFerran BW, Bodill H, et al. Similar effects of α- and β-SNAP on Ca2+-regulated exocytosis. FEBS Lett, 1996,393(2-3):185-8
3 郑芳,吴群,姜小丹,等. NK92 细胞的胞吐效应与 SNAP-23 蛋白转位相关. 中华微生物学和免疫学杂志,2005,2(25) :85-88
4 Borowsky B, Hoffman BJ. Neurotransmitter transporters: molecular biology, function, and regulation. Int Rev Neurobiol, 1995,38:139-99
5 Bonisch H, Bruss M. The noradrenaline transporter of the neuronal plasma membrane. Ann N Y Acad Sci, 1994,733:193-202
6 曹君利,丁海雷,杨国栋,等. 囊泡胞吐机制及与其相关的神经元可塑性. 生命科学,2003,15:283-288
7 Brose N, Petrenko AG, Sudhof TC, et al. Synaptotagmin: a calcium sensor on the synaptic vesicle surface. Science, 1992,256:1021-5
8 Adolfsen B, Littleton JT. Genetic and molecular analysis of the synaptotagmin family. Cell Mol Life Sci, 2001,58:393-402
9 Lonart G. RIM1: an edge for presynaptic plasticity. Trends Neurosci, 2002,25:329-32
10 Rizo J, Sudhof TC. Snares and Munc18 in synaptic vesicle fusion. Nat Rev Neurosci, 2002,3:641-53
11 Betz A, Thakur P, Junge HJ, et al. Functional interaction of the active zone proteins Munc13-1 and RIM1 in synaptic vesicle priming. Neuron, 2001,30:183-96
12 Morgan A, Dimaline R, Burgoyne RD. The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. J Biol Chem, 1994,269:29347-50
13 Griff IC, Schekman R, Rothman JE, et al. The yeast SEC17 gene product is functionally equivalent to mammalian α-SNAP protein. J Biol Chem, 1992,267(17):12106-15
14 Peter F, Wong SH, Subramaniam VN, et al. Α-SNAP but not γ-SNAP is required for ER-Golgi transport after vesicle budding and the Rab1-requiring step but before the EGTA-sensitive step. J Cell Sci, 1998,111 ( Pt 17):2625-33
15 Reuveny E, Narahashi T. Two types of high voltage-activated calcium channels in SH-SY5Y human neuroblastoma cells. Brain Res, 1993,603:64-73
16 Purkiss JR, Friis LM, Doward S, et al. Clostridium botulinum neurotoxinsact with a wide range of potencies on SH-SY5Y human neuroblastoma cells. Neurotoxicology, 2001,22:447-53.
17 Paddock SW. Principles and practices of laser scanning confocal microscopy. Mol Biotechnol ,2000,16(2): 127-149
18 Kreft M, Stenovec M, Zorec R. Focus-drift correction in time-lapse confocal imaging. Ann NY Acad Sci ,2005,1048(6):321-330
19 Collazo A, Bricaud O, Desai K. Use of confocal microscopy in comparative studies of vertebrate morphology. Methods Enzymol, 2005,395:521-543
20 Seyedi N, Mackins CJ, Machida T, et al. Histamine H3-receptor-induced attenuation of norepinephrine exocytosis: a decreased protein kinase a activity mediates a reduction in intracellular calcium. J Pharmacol Exp Ther, 2005,312:272-80
21 Wang Z, Sadee W. Tolerance to morphine at the mu-opioid receptor differentially induced by cAMP-dependent protein kinase activation and morphine. Eur J Pharmacol,2000,389(2-3):165-71
22 Nestler EJ, Alreja M, Aghajanian GK. Molecular and cellular mechanisms of opiate action: studies in the rat locus coeruleus. Brain Res Bull, 1994,35(5-6):521-8
23 Nestler EJ, Tallman JF. Chronic morphine treatment increases cyclic AMP-dependent protein kinase activity in the rat locus coeruleus. Mol Pharmacol,1988,33(2):127-32
24 Eriksson PS, Carlsson B, Isaksson OG, et al. Altered amounts of G-protein mRNA and cAMP accumulation after long-term opioid receptor stimulation of neurons in primary culture from the rat cerebral cortex. Brain Res Mol Brain Res, 1992,14(4):317-25
25 Duman RS, Tallman JF, Nestler EJ. Acute and chronic opiate-regulation of adenylate cyclase in brain: specific effects in locus coeruleus. J Pharmacol Exp Ther,1988,246(3):1033-9
26 Nestler EJ. Molecular basis of long-term plasticity underlying addiction. Nat Rev Neurosci,2001,2(2):119-28
27 Llinas R, Sugimori M, Silver RB. Microdomains of high calcium concentration in a presynaptic terminal. Science, 1992,256:677-9.
28 Zucker RS. Calcium and transmitter release. J Physiol Paris, 1993,87:25-36
29 Bai J, Wang CT, Richards DA, et al. Fusion pore dynamics are regulated by synaptotagmin*t-SNARE interactions. Neuron, 2004,41:929-42
30 Stenbeck G. Soluble NSF-attachment proteins. Int J Biochem Cell Biol, 1998,30:573-7
31 Nishiki T, Nihonmatsu I, Tsuhara Y, et al. Distribution of soluble N-ethylmaleimide fusion protein attachment proteins (SNAPs) in the rat nervous system. Neuroscience, 2001,107:363-71
32 Sollner T. SNAREs and targeted membrane fusion. FEBS Lett, 1995,369:80-3
33 Woodman PG. The roles of NSF, SNAPs and SNAREs during membrane fusion. Biochim Biophys Acta, 1997,1357:155-72
34 Sheng ZH, Yokoyama CT, Catterall WA. Interaction of the synprint site of N-type Ca2+ channels with the C2B domain of synaptotagmin I. Proc Natl Acad Sci U S A, 1997,94:5405-10
35 Rettig J, Sheng ZH, Kim DK, et al. Isoform-specific interaction of the alpha1A subunits of brain Ca2+ channels with the presynaptic proteins syntaxin and SNAP-25. Proc Natl Acad Sci U S A, 1996,93:7363-8
36 Sheng ZH, Rettig J, Cook T, et al. Calcium-dependent interaction of N-type calcium channels with the synaptic core complex. Nature, 1996,379:451-4
37 Mochida S, Sheng ZH, Baker C, et al. Inhibition of neurotransmission by peptides containing the synaptic protein interaction site of N-type Ca2+ channels. Neuron, 1996,17:781-8
38 杜宝国,张瑞玲,李昌琪,等.吗啡依赖大鼠伏隔核区单胺转运体表达的时点变化.中国药物依赖性杂志,2006,15:358-362
39 Xu NJ, Bao L, Fan HP, et al. Morphine withdrawal increases glutamate uptake and surface expression of glutamate transporter GLT1 at hippocampal synapses. J Neurosci, 2003,23:4775-84
40 Gandhi SP, Stevens CF. Three modes of synaptic vesicular recycling revealed by single-vesicle imaging. Nature, 2003,423:607-13
1 Nishiki T, Nihonmatsu I, Tsuhara Y, et al. Distribution of soluble N-ethylmaleimide fusion protein attachment proteins (SNAPs) in the rat nervous system. Neuroscience, 2001,107:363-71
2 Whiteheart SW, Griff IC, Brunner M, et al. SNAP family of NSF attachment proteins includes a brain-specific isoform. Nature, 1993,362:353-5
3 Stenbeck G. Soluble NSF-attachment proteins. Int J Biochem Cell Biol, 1998,30:573-7
4 Kato K. A Collection of cDNA Clones with Specific Expression Patterns in Mouse Brain. Eur J Neurosci, 1990,2:704-711
5 Rothman JE, Warren G. Implications of the SNARE hypothesis for intracellular membrane topology and dynamics. Curr Biol, 1994,4:220-33
6 Cho SJ, Kelly M, Rognlien KT, et al. SNAREs in opposing bilayers interact in a circular array to form conducting pores. Biophys J, 2002,83:2522-7
7 Cho WJ, Jeremic A, Jena BP. Size of supramolecular SNARE complex: membrane-directed self-assembly. J Am Chem Soc, 2005,127:10156-7
8 Sutton RB, Fasshauer D, Jahn R, et al. Crystal structure of a SNARE complex involved in synaptic exocytosis at 2.4 A resolution. Nature, 1998,395:347-53
9 Hayashi T, McMahon H, Yamasaki S, et al. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly. EMBO J, 1994,13:5051-61
10 Yang B, Gonzalez L Jr, Prekeris R, et al. SNARE interactions are not selective. Implications for membrane fusion specificity. J Biol Chem, 1999,274:5649-53
11 Hanson PI, Heuser JE, Jahn R. Neurotransmitter release - four years of SNARE complexes. Curr Opin Neurobiol, 1997,7:310-5
12 Brunger AT. Structure and function of SNARE and SNARE-interactingproteins. Q Rev Biophys, 2005,38:1-47
13 Jahn R, Scheller RH. SNAREs--engines for membrane fusion. Nat Rev Mol Cell Biol, 2006,7:631-43
14 Sudhof TC. The synaptic vesicle cycle. Annu Rev Neurosci, 2004,27:509-47
15 Fasshauer D, Margittai M. A transient N-terminal interaction of SNAP-25 and syntaxin nucleates SNARE assembly. J Biol Chem, 2004,279:7613-21
16 Fasshauer D, Otto H, Eliason WK, et al. Structural changes are associated with soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor complex formation. J Biol Chem, 1997,272:28036-41
17 Sorensen JB, Wiederhold K, Muller EM, et al. Sequential N- to C-terminal SNARE complex assembly drives priming and fusion of secretory vesicles. EMBO J, 2006,25:955-66
18 Weber T, Zemelman BV, McNew JA, et al. SNAREpins: minimal machinery for membrane fusion. Cell, 1998,92:759-72
19 McNew JA, Weber T, Parlati F, et al. Close is not enough: SNARE-dependent membrane fusion requires an active mechanism that transduces force to membrane anchors. J Cell Biol, 2000,150:105-17
20 Hu C, Ahmed M, Melia TJ, et al. Fusion of cells by flipped SNAREs. Science, 2003,300:1745-9
21 Fix M, Melia TJ, Jaiswal JK, et al. Imaging single membrane fusion events mediated by SNARE proteins. Proc Natl Acad Sci U S A, 2004,101:7311-6
22 Sollner T, Bennett MK, Whiteheart SW, et al. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell, 1993,75:409-18
23 Clary DO, Griff IC, Rothman JE. SNAPs, a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast. Cell, 1990,61:709-21
24 Wilson DW, Whiteheart SW, Wiedmann M, et al. A multisubunit particle implicated in membrane fusion. J Cell Biol, 1992,117:531-8
25 Morgan A, Dimaline R, Burgoyne RD. The ATPase activity of N-ethylmaleimide-sensitive fusion protein (NSF) is regulated by soluble NSF attachment proteins. J Biol Chem, 1994,269:29347-50
26 Matveeva E, Whiteheart SW. The effects of SNAP/SNARE complexes on the ATPase of NSF. FEBS Lett, 1998,435:211-4
27 Hayashi T, Yamasaki S, Nauenburg S, et al. Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. EMBO J, 1995,14:2317-25
28 McMahon HT, Sudhof TC. Synaptic core complex of synaptobrevin, syntaxin, and SNAP-25 forms high affinity alpha-SNAP binding site. J Biol Chem, 1995,270:2213-7
29 Tagaya M, Wilson DW, Brunner M, et al. Domain structure of an N-ethylmaleimide-sensitive fusion protein involved in vesicular transport. J Biol Chem, 1993,268:2662-6
30 Whiteheart SW, Brunner M, Wilson DW, et al. Soluble N-ethylmaleimide-sensitive fusion attachment proteins (SNAPs) bind to a multi-SNAP receptor complex in Golgi membranes. J Biol Chem, 1992,267:12239-43
31 Wimmer C, Hohl TM, Hughes CA, et al. Molecular mass, stoichiometry, and assembly of 20 S particles. J Biol Chem, 2001,276:29091-7
32 Hanson PI, Otto H, Barton N, et al. The N-ethylmaleimide-sensitive fusion protein and alpha-SNAP induce a conformational change in syntaxin. J Biol Chem, 1995,270:16955-61
33 Barnard RJ, Morgan A, Burgoyne RD. Domains of alpha-SNAP required for the stimulation of exocytosis and for N-ethylmalemide-sensitive fusion protein (NSF) binding and activation. Mol Biol Cell, 1996,7:693-701
34 Barnard RJ, Morgan A, Burgoyne RD. Stimulation of NSF ATPase activity by alpha-SNAP is required for SNARE complex disassembly andexocytosis. J Cell Biol, 1997,139:875-83
35 Tani K, Shibata M, Kawase K, et al. Mapping of functional domains of gamma-SNAP. J Biol Chem, 2003,278:13531-8
36 Puschel AW, O'Connor V, Betz H. The N-ethylmaleimide-sensitive fusion protein (NSF) is preferentially expressed in the nervous system. FEBS Lett, 1994,347:55-8
37 DeBello WM, O'Connor V, Dresbach T, et al. SNAP-mediated protein-protein interactions essential for neurotransmitter release. Nature JT - Nature, 1995,373:626-30
38 Hirling H, Scheller RH. Phosphorylation of synaptic vesicle proteins: modulation of the alpha SNAP interaction with the core complex. Proc Natl Acad Sci U S A, 1996,93:11945-9
39 DiAntonio A, Parfitt KD, Schwarz TL. Synaptic transmission persists in synaptotagmin mutants of Drosophila. Cell, 1993,73:1281-90
40 Littleton JT, Stern M, Schulze K, et al. Mutational analysis of Drosophila synaptotagmin demonstrates its essential role in Ca(2+)-activated neurotransmitter release. Cell, 1993,74:1125-34
41 DiAntonio A, Schwarz TL. The effect on synaptic physiology of synaptotagmin mutations in Drosophila. Neuron, 1994,12:909-20
42 Broadie K, Bellen HJ, DiAntonio A, et al. Absence of synaptotagmin disrupts excitation-secretion coupling during synaptic transmission. Proc Natl Acad Sci U S A, 1994,91:10727-31
43 Griff IC, Schekman R, Rothman JE, et al. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein. J Biol Chem JT, 1992,267:12106-15
44 Schiavo G, Gmachl MJ, Stenbeck G, et al. A possible docking and fusion particle for synaptic transmission. Nature, 1995,378:733-6
45 Martin HG, Henley JM, Meyer G. Novel putative targets of N-ethylmaleimide sensitive fusion protein (NSF) and alpha/beta soluble NSF attachment proteins (SNAPs) include the Pak-binding nucleotide exchange factor betaPIX. J Cell Biochem, 2006,99:1203 -15
46 Peter F, Wong SH, Subramaniam VN, et al. Alpha-SNAP but not gamma-SNAP is required for ER-Golgi transport after vesicle budding and the Rab1-requiring step but before the EGTA-sensitive step. J Cell Sci , 1998,111 ( Pt 17):2625-33
47 Nagamatsu S, Watanabe T, Nakamichi Y, et al. alpha-soluble N-ethylmaleimide-sensitive factor attachment protein is expressed in pancreatic beta cells and functions in insulin but not gamma-aminobutyric acid secretion. J Biol Chem,1999,274:8053 -60
48 Morgan A, Burgoyne RD. A role for soluble NSF attachment proteins (SNAPs) in regulated exocytosis in adrenal chromaffin cells. EMBO J, 1995,14:232-9
49 Abonyo BO, Wang P, Narasaraju TA, et al. Characterization of alpha-soluble N-ethylmaleimide-sensitive fusion attachment protein in alveolar type II cells: implications in lung surfactant secretion. Am J Respir Cell Mol Biol, 2003,29:273-82
50 Baur T, Ramadan K, Schlundt A, et al. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts. J Cell Sci, 2007,120:2895-903
51 Echard A, Hickson GR, Foley E, et al. Terminal cytokinesis events uncovered after an RNAi screen. Curr Biol, 2004,14:1685-93
52 Sudlow AW, McFerran BW, Bodill H, et al. Similar effects of alpha- and beta-SNAP on Ca2+-regulated exocytosis. FEBS Lett, 1996,393:185-8
53 Xu J, Xu Y, Ellis-Davies GC, et al. Differential regulation of exocytosis by alpha- and beta-SNAPs. J Neurosci, 2002,22:53-61
54 Andreeva AV, Kutuzov MA, Voyno-Yasenetskaya TA. A ubiquitous membrane fusion protein alpha SNAP: a potential therapeutic target for cancer, diabetes and neurological disorders? Expert Opin Ther Targets, 2006,10:723-33
55 Yoo BC, Cairns N, Fountoulakis M, et al. Synaptosomal proteins, beta-soluble N-ethylmaleimide-sensitive factor attachment protein(beta-SNAP), gamma-SNAP and synaptotagmin I in brain of patients with Down syndrome and Alzheimer's disease. Dement Geriatr Cogn Disord, 2001,12:219-25
56 Sultana R, Boyd-Kimball D, Poon HF, et al. Redox proteomics identification of oxidized proteins in Alzheimer's disease hippocampus and cerebellum: an approach to understand pathological and biochemical alterations in AD. Neurobiol Aging, 2006,27:1564-76
57 Weller AE, Dahl JP, Lohoff FW, et al. Analysis of variations in the NAPG gene on chromosome 18p11 in bipolar disorder. Psychiatr Genet, 2006,16:3-8
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |