大鼠前扣带回皮质5-HT_(1A,1B,2A,2C)受体亚型蛋白表达的细胞定位
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摘要
目的:观察5-HT_(1A、1B)和5HT_(2A、2C)等受体在大鼠前扣带回皮质(anterior cingulated cortex,ACC)的分布及细胞定位。方法:将20只成年雄性Sprague-Dawley大鼠平均分成四组,每组5只分别进行四种亚型5-HT的免疫组织化学反应。动物经过灌注固定、脑冠状切片(25μm)后,以卵白素-生物素复合物法(ABC法)对切片进行染色、显微镜观察。结果:四种5-HT受体亚型在ACC都有分布。ACC内II-VI层均有5-HT_(1A)免疫阳性反应神经元的胞体和树突。5-HT_(1B)受体亚型定位在神经元胞体上,表达较弱。I-III层5-HT_(2A)免疫阳性反应较强,主要为标记的锥体神经元树突(Cg1区III层观察到部分阳性反应的神经元胞体);而V-VI层可见到明显的免疫阳性神经元胞体。5-HT2C受体定位于ACC之II-VI层神经元胞体上。结论:5-HT_(1A)、5-HT_(1B)、5-HT_(2A)和5-HT_(2C)等不同受体亚型在ACC内细胞上的定位有一定差异,提示它们介导5-HT对ACC神经元发生调节反应的细胞作用部位有选择性,因之它们介导的5-HT对ACC神经元的作用效果也可能不同。
Objective: To examine the laminar distribution and cellular localization of 5-hydroxytryptamine( 5-HT)receptor subtype proteins of 5-HT_(1A,1B) and 5-HT_(2A,2C) in the anterior cingulate cortex( ACC) of rat. Methods: A total of twenty normal male Sprague-Dawley( SD) rats were used,which were equally divided into four groups of five rats each,immunohistochemistry for the four subtypes of 5-HT receptors,respectively. Animals were fixed via perfusion and brains were cut into coronal slices in 25 μm thickness. Sections were immunostained with Avidin-Biotin Complex( ABC) method before observation with light microscopy. Results: Immunoreactivities of all four 5-HT receptor subtypes were detected throughout the whole rostrocaudal extent of ACC. Laminae II through to VI of ACC contained 5-HT_(1A) immunopositive cell bodies and dendrites; 5-HT_(1B) receptor was localized to cell bodies as shown by relatively weak immunoreactivity. High intensity of 5-HT_(2A) immunolabeling was present in laminae I to III of ACC,appearing as labeled dendrites of pyramidal neurons with the presence of additional labeled cell bodies in layer III of area 1(Cg1); while laminae V and VI witnessed 5-HT_(2A) immunopositive neuronal cell bodies. 5-HT_(2C) receptor was localized to neuronal cell bodies of laminae II through to VI. Conclusion: Laminar distribution and cellular localization with respect to 5-HT_(1 A),5-HT_(1B),5-HT_(2A) and 5-HT_(2C) receptors showed distinguishable difference,suggesting the selectivity of the individual receptors in cellular compartments for mediation of 5-HT actions,which may yield different effects of 5-HT on the resulting activity of ACC neurons.
Objective: To examine the laminar distribution and cellular localization of 5-hydroxytryptamine( 5-HT)receptor subtype proteins of 5-HT_(1A,1B) and 5-HT_(2A,2C) in the anterior cingulate cortex( ACC) of rat. Methods: A total of twenty normal male Sprague-Dawley( SD) rats were used,which were equally divided into four groups of five rats each,immunohistochemistry for the four subtypes of 5-HT receptors,respectively. Animals were fixed via perfusion and brains were cut into coronal slices in 25 μm thickness. Sections were immunostained with Avidin-Biotin Complex( ABC) method before observation with light microscopy. Results: Immunoreactivities of all four 5-HT receptor subtypes were detected throughout the whole rostrocaudal extent of ACC. Laminae II through to VI of ACC contained 5-HT_(1A) immunopositive cell bodies and dendrites; 5-HT_(1B) receptor was localized to cell bodies as shown by relatively weak immunoreactivity. High intensity of 5-HT_(2A) immunolabeling was present in laminae I to III of ACC,appearing as labeled dendrites of pyramidal neurons with the presence of additional labeled cell bodies in layer III of area 1(Cg1); while laminae V and VI witnessed 5-HT_(2A) immunopositive neuronal cell bodies. 5-HT_(2C) receptor was localized to neuronal cell bodies of laminae II through to VI. Conclusion: Laminar distribution and cellular localization with respect to 5-HT_(1 A),5-HT_(1B),5-HT_(2A) and 5-HT_(2C) receptors showed distinguishable difference,suggesting the selectivity of the individual receptors in cellular compartments for mediation of 5-HT actions,which may yield different effects of 5-HT on the resulting activity of ACC neurons.
引文
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[23]Puig MV,Gulledge AT.Serotonin and prefrontal cortex function:neurons,networks,and circuits[J].Mol Neurobiol,2011,44:449-464.
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[25]Chiba H,Mitsui T,Kitta T,et al.The role of serotonergic mechanism in the rat prefrontal cortex for controlling the micturition reflex:an in vivo microdialysis study[J].Neurourol Urodyn,2015,1-6.
[26]Ding S,Li L,Zhou FM.Robust presynaptic serotonin 5-HT(1B)receptor inhibition of the striatonigral output and its sensitization by chronic fluoxetine treatment[J].J Neurophysiol,2015,113:3397-3409.
[27]Potenza MN,Walderhaug E,Henry S,et al.Serotonin 1B receptor imaging in pathological gambling[J].World J Biol Psychiatry,2013,14:139-145.
[28]Matuskey D,Bhagwagar Z,Planeta B,et al.Reductions in brain5-HT1B receptor availability in primarily cocaine-dependent humans[J].Biol Psychiatry,2014,76:816-822.
[29]Anastasio NC,Lanfranco MF,Bubar MJ,et al.Serotonin 5-HT2Creceptor protein expression is enriched in synaptosomal and postsynaptic compartments of rat cortex[J].J Neurochem,2010,113:1504-1515.
[30]Aghajanian GK,Marek GJ.Serotonin induces excitatory postsynaptic potentials inapical dendrites of neocortical pyramidal cells[J].Neuropharmacology,1997,36:589-599.
[31]Beique JC,Campbell B,Perring P,et al.Serotonergic regulation of membrane potential in developing rat prefrontal cortex:coordinated expression of 5-hydroxytryptamine(5-HT)1A,5-HT2A,and 5-HT7 receptors[J].J Neurosci,2004,24:4807-4817.
[32]Leiser SC,Li Y,Pehrson AL,et al.Serotonergic regulation of prefrontal cortical circuitries involved in cognitive processing:A review of individual 5-HT receptor mechanisms and concerted effects of 5-HT receptors exemplified by the multimodal antidepressant vortioxetine[J].ACS Chem Neurosci,2015,6:970-986.
[33]Aghajanian GK,Marek GJ.Serotonin,via 5-HT2A receptors,increases EPSCs in layer V pyramidal cells of prefrontal cortex by an asynchronous mode of glutamate release[J].Brain Res,1999,825:161-171.
[34]Arvanov VL,Wang RY.M100907,a selective 5-HT2A receptor antagonist and a potential antipsychotic drug,facilitates N-methylD-aspartate-receptor mediated neurotransmission in the rat medial prefrontal cortical neurons in vitro[J].Neuropsychopharmacology,1998,18:197-209.
[35]Yuen EY,Jiang Q,Chen P,et al.Serotonin 5-HT1A receptors regulate NMDA receptor channels through a microtubule-dependent mechanism[J].J Neurosci,2005,25:5488-5501.
[36]Yuen EY,Jiang Q,Chen P,et al.Activation of 5-HT2A/C receptors counteracts 5-HT1A regulation of n-methyl-D-aspartate receptor channels in pyramidal neurons ofprefrontal cortex[J].J Biol Chem,2008,283:17194-17204.
[2]Hoyer D,Hannon JP,Martin GR.Molecular,pharmacological and functional diversity of 5-HT receptors[J].Pharmacol,Biochem Behav,2002,71:533-554.
[3]Hannon J,Hoyer D.Molecular biology of 5-HT receptors[J].Behav Brain Res,2008,195:198-213.
[4]Hoyer D,Clarke DE,Fozard JR,et al.International union of pharmacology classification of receptors for 5-hydroxytryptamine(serotonin)[J].Pharmacol Rev,1994,46:157-203.
[5]Hadland KA,Rushworth MFS,Gaffan D,et al.The effect of cingulate lesions on social behaviour and emotion[J].Neuropsychologia,2003,41:919-931.
[6]Kozlovskiy SA,Vartanov AV,Nikonova EY,et al.The cingulate cortex and human memory processes[J].Psychol Russ State Art,2012,5:231-243.
[7]Kozlovskiy S,Vartanov A,Pyasik M,et al.Anatomical characteristics of cingulate cortex and neuropsychological memory tests performance[J].Proc Social Behav Sci,2013,86:128-133.
[8]Vogt BA,Finch DM,Olson CR.Functional heterogeneity in cingulate cortex:the anterior executive and posterior evaluative regions[J].Cerebral Cortex,1992,2:435-443.
[9]Devinsky O,Morrell MJ,Vogt BA.Contributions of anterior cingulate cortex to behaviour[J].Brain,1995,118:279-306.
[10]Bush G,Luu P,Posner MI.Cognitive and emotionalinfluences in anteriorcingulate cortex[J].Trends Cogn Sci,2000,4:215-222.
[11]Uylings HBM,Groenewegen HJ,Kolb B.Do rats have a prefrontal cortex?[J].Behav Brain Res,2003,146:3-17.
[12]Vertes RP.A PHA-L analysis of ascendingprojections of the dorsal raphe nucleusin the rat[J].J Comp Neurol,1991,313:643-668.
[13]Linley SB,Hoover WB,Vertes RP.Pattern of distribution of serotonergic fibers to the orbitomedial andinsular cortex in the rat[J].JChem Neuroanat,2013,45:48-49.
[14]Barnes NM,Sharp T.A review of central 5-HT receptors and their function[J].Neuropharmacology,1999,38:1083-1152.
[15]Wright DE,Seroogy KB,Lundgren KH,et al,Comparative localization of serotonin1A,1C,and 2 receptor subtype mRNAs in rat brain[J].J Comp Neurol,1995,351:357-373.
[16]Aznar S,Qian Z,Shah R,et al.The 5-HT1A serotonin receptor is located on calbindin-and parvalbumin-containing neurons in the rat brain[J].Brain Res,2003,959:58-67.
[17]Sari Y.Serotonin1B receptors:from protein to physiological function and behavior.Neurosci Biobehav Rev,2004,28:565-582.
[18]Kia HK,Miquel MC,Brisorgueil MJ,et al.Immunocytochemical localization of serotonin1A receptors in the rat central nervous system[J].J Comp Neurol,1996,365:289-305.
[19]Cruz DA,Eggan SM,Azmitia EC,et al.Serotonin 1A receptors at the axon initial segment of prefrontal pyramidal neurons in schizophrenia[J].Am J Psychiatry,2004.161:p.739-742.
[20]Llado PL,Santana N,Ghisi V,et al.5-HT1A receptor agonists enhance pyramidal cell firing in prefrontal cortex through a preferential action on GABA interneurons[J].Cereb Cortex,2012 22:1487-1497.
[21]Stephens EK,Avesar D,Gulledge AT.Activity-dependent serotonergic excitation of callosal projection neurons in the mouse prefrontal cortex[J].Front Neural Circuits,2014,8:97.
[22]Villalobos C,Beique JC,Gingrich JA,et al.Serotonergic regulation of calcium-activated potassium currents in rodent prefrontal cortex[J].Eur J Neurosci,2005,22:1120-1126.
[23]Puig MV,Gulledge AT.Serotonin and prefrontal cortex function:neurons,networks,and circuits[J].Mol Neurobiol,2011,44:449-464.
[24]Amargos BM,Bortolozzi A,Puig MA,et al.Co-expression and in vivo interaction of serotonin 1A and serotonin 2A receptors in pyramidal neurons of prefrontal cortex[J].Cereb Cortex,2004,14:281-299.
[25]Chiba H,Mitsui T,Kitta T,et al.The role of serotonergic mechanism in the rat prefrontal cortex for controlling the micturition reflex:an in vivo microdialysis study[J].Neurourol Urodyn,2015,1-6.
[26]Ding S,Li L,Zhou FM.Robust presynaptic serotonin 5-HT(1B)receptor inhibition of the striatonigral output and its sensitization by chronic fluoxetine treatment[J].J Neurophysiol,2015,113:3397-3409.
[27]Potenza MN,Walderhaug E,Henry S,et al.Serotonin 1B receptor imaging in pathological gambling[J].World J Biol Psychiatry,2013,14:139-145.
[28]Matuskey D,Bhagwagar Z,Planeta B,et al.Reductions in brain5-HT1B receptor availability in primarily cocaine-dependent humans[J].Biol Psychiatry,2014,76:816-822.
[29]Anastasio NC,Lanfranco MF,Bubar MJ,et al.Serotonin 5-HT2Creceptor protein expression is enriched in synaptosomal and postsynaptic compartments of rat cortex[J].J Neurochem,2010,113:1504-1515.
[30]Aghajanian GK,Marek GJ.Serotonin induces excitatory postsynaptic potentials inapical dendrites of neocortical pyramidal cells[J].Neuropharmacology,1997,36:589-599.
[31]Beique JC,Campbell B,Perring P,et al.Serotonergic regulation of membrane potential in developing rat prefrontal cortex:coordinated expression of 5-hydroxytryptamine(5-HT)1A,5-HT2A,and 5-HT7 receptors[J].J Neurosci,2004,24:4807-4817.
[32]Leiser SC,Li Y,Pehrson AL,et al.Serotonergic regulation of prefrontal cortical circuitries involved in cognitive processing:A review of individual 5-HT receptor mechanisms and concerted effects of 5-HT receptors exemplified by the multimodal antidepressant vortioxetine[J].ACS Chem Neurosci,2015,6:970-986.
[33]Aghajanian GK,Marek GJ.Serotonin,via 5-HT2A receptors,increases EPSCs in layer V pyramidal cells of prefrontal cortex by an asynchronous mode of glutamate release[J].Brain Res,1999,825:161-171.
[34]Arvanov VL,Wang RY.M100907,a selective 5-HT2A receptor antagonist and a potential antipsychotic drug,facilitates N-methylD-aspartate-receptor mediated neurotransmission in the rat medial prefrontal cortical neurons in vitro[J].Neuropsychopharmacology,1998,18:197-209.
[35]Yuen EY,Jiang Q,Chen P,et al.Serotonin 5-HT1A receptors regulate NMDA receptor channels through a microtubule-dependent mechanism[J].J Neurosci,2005,25:5488-5501.
[36]Yuen EY,Jiang Q,Chen P,et al.Activation of 5-HT2A/C receptors counteracts 5-HT1A regulation of n-methyl-D-aspartate receptor channels in pyramidal neurons ofprefrontal cortex[J].J Biol Chem,2008,283:17194-17204.