β2-nAChR促进小鼠海马CA1和CA3锥体神经元GABA_A受体的功能成熟
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摘要
目的探讨β2-烟碱型乙酰胆碱受体(β2-n ACh R)在海马CA1和CA3锥体神经元的A型γ-氨基丁酸受体(GABA_A-R)发育中的作用。方法应用β2-n ACh R基因敲除小鼠(β2-KO组)制备急性分离的海马CA1和CA3锥体神经元,应用穿孔膜片钳记录技术记录GABA_A-R选择性激动剂蝇蕈醇在CA1和CA3锥体神经元诱导的GABA电流,测试其平衡电位(E_(Mus))和动力学指标,并与野生型小鼠(WT组)进行比较。结果β2-KO组小鼠(n=4)CA1锥体神经元(n=7)的E_(Mus)为-31.7±3.5 m V,与WT组相比向去极化偏移(P<0.05);CA3锥体神经元(n=4)的E_(Mus)为-16.1±4.6 m V,同样较WT组偏向去极化方向(P<0.01);与WT组小鼠不同,β2-KO组小鼠CA3和CA1神经元的E_(Mus)差异有统计学意义(P<0.05)。β2-KO组小鼠CA1和CA3神经元上都显示GABA_A-R的失敏显著减慢,衰减时间分别为2.2±0.2 s、3.2±0.1 s(WT组为1.6±0.1 s、2.3±0.1 s,P<0.05或P<0.01)。结论含β2的n ACh R可能参与促进小鼠海马CA1和CA3锥体细胞中GABA_A-R的功能成熟。
Objective To explore the role of β2-nicotinic acetylcholine receptor(β2-n ACh R) in the development of γ-aminobutyric acid A type receptors(GABA_A-Rs) in hippocampal CA1 and CA3 pyramidal neurons of mice. Methods The hippocampal CA1 and CA3 pyramidal neurons were acutely isolated from β2-n ACh R gene knockout(β2-KO group) mice.GABA currents in CA1 and CA3 pyramidal neurons were induced with the selective GABA_A-R agonist muscimol and recorded using perforated patch-clamp recording technique. The GABA currents of CA1 and CA3 pyramidal neurons were tested for their equilibrium potentials(E_(Mus)s) and kinetic parameters and were compared with the measurements in wild-type mice(WT group). Results The mean E_(Mus) of CA1 neurons(n=7) of β2-KO mice(n=4) was-31.7±3.5 m V, showing an obvious depolarizing shift compared with the WT mice(P<0.05); the mean E_(Mus)of CA3 neurons(n=4) was-16.1±4.6 m V, also showing a depolarizing shift(P<0.01). The difference in the E_(Mus)s between CA3 and CA1 neurons in β2-KO mice, but not in WT mice, was significant(P<0.05). The GABA_A-R desensitization was significantly slowed down in both CA1 and CA3 neurons of β2-KO mice, with decay time of 2.2±0.2 s and 3.2±0.1 s, respectively, significantly longer than those in WT mice(1.6±0.1 s and 2.3±0.1 s, respectively; P<0.05). Conclusion β2-containing n ACh Rs may promote the functional maturation of GABA_A-R in CA1 and CA3 pyramidal cells in mouse hippocampus.
Objective To explore the role of β2-nicotinic acetylcholine receptor(β2-n ACh R) in the development of γ-aminobutyric acid A type receptors(GABA_A-Rs) in hippocampal CA1 and CA3 pyramidal neurons of mice. Methods The hippocampal CA1 and CA3 pyramidal neurons were acutely isolated from β2-n ACh R gene knockout(β2-KO group) mice.GABA currents in CA1 and CA3 pyramidal neurons were induced with the selective GABA_A-R agonist muscimol and recorded using perforated patch-clamp recording technique. The GABA currents of CA1 and CA3 pyramidal neurons were tested for their equilibrium potentials(E_(Mus)s) and kinetic parameters and were compared with the measurements in wild-type mice(WT group). Results The mean E_(Mus) of CA1 neurons(n=7) of β2-KO mice(n=4) was-31.7±3.5 m V, showing an obvious depolarizing shift compared with the WT mice(P<0.05); the mean E_(Mus)of CA3 neurons(n=4) was-16.1±4.6 m V, also showing a depolarizing shift(P<0.01). The difference in the E_(Mus)s between CA3 and CA1 neurons in β2-KO mice, but not in WT mice, was significant(P<0.05). The GABA_A-R desensitization was significantly slowed down in both CA1 and CA3 neurons of β2-KO mice, with decay time of 2.2±0.2 s and 3.2±0.1 s, respectively, significantly longer than those in WT mice(1.6±0.1 s and 2.3±0.1 s, respectively; P<0.05). Conclusion β2-containing n ACh Rs may promote the functional maturation of GABA_A-R in CA1 and CA3 pyramidal cells in mouse hippocampus.
引文
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[2]Zheng F,Khanna S.Intra-hippocampal tonic inhibition influences formalin pain-induced pyramidal cell suppression,but not excitation in dorsal field CA1 of rat[J].Brain Res Bull,2008,77(6):374-81.
[3]Rambousek L,Kleteckova L,Kubesova A,et al.Rat intrahippocampal NMDA infusion induces cell-specific damage and changes in expression of NMDA and GABA(A)receptor subunits[J].Neuropharmacology,2016,105(13):594-606.
[4]Sibbe M,Kulik A.GABAergic regulation of adult hippocampal neurogenesis[J].Mol Neurobiol,2017,54(7):5497-510.
[5]Lee V,Mackenzie G,Hooper A,et al.Reduced tonic inhibition in the dentate gyrus contributes to chronic stress-induced impairments in learning and memory[J].Hippocampus,2016,26(10):1276-90.
[6]Hurst R,Rollema H,Bertrand D.Nicotinic acetylcholine receptors:From basic science to therapeutics[J].Pharmacol Ther,2013,137(1):22-54.
[7]Kaneko N,Okano H,Sawamoto K.Role of the cholinergic system in regulating survival of newborn neurons in the adult mouse dentate gyrus and olfactory bulb[J].Genes Cells,2006,11(10):1145-59.
[8]Ide Y,Fujiyama F,Okamoto-Furuta K,et al.Rapid integration of young newborn dentate gyrus granule cells in the adult hippocampal circuitry[J].Eur J Neurosci,2008,28(12):2381-92.
[9]King MD,Long T,Andersen T,et al.Genetic algorithm managed peptide mutant screening:optimizing peptide ligands for targeted receptor binding[J].J Chem Inf Model,2016,56(12):2378-87.
[10]Lange-Asschenfeldt C,Sch?ble S,Suvorava T,et al.Effects of varenicline on alpha4-containing nicotinic acetylcholine receptor expression and cognitive performance in mice[J].Neuropharmacology,2016,107(21):100-10.
[11]Campbell NR,Fernandes CC,Halff AW,et al.Endogenous signaling through alpha7-containing nicotinic receptors promotes maturation and integration of adult-born neurons in the hippocampus[J].J Neurosci,2010,30(26):8734-44.
[12]Liu ZP,Zhang JM,Berg DK.Role of endogenous nicotinic signaling in guiding neuronal development[J].Biochem Pharmacol,2007,74(8):1112-9.
[13]Yalcin I,Charlet A,Cordero-Erausquin M,et al.Nociceptive thresholds are controlled through spinal beta(2)-subunit-containing nicotinic acetylcholine receptors[J].Pain,2011,152(9):2131-7.
[14]Pereira Silva JD,Campos DV,Nogueira-Bechara FM,et al.Altered release and uptake of gamma-aminobutyric acid in the cerebellum of dystrophin-deficient mice[J].Neurochem Int,2018,85(6):1-14.
[15]Nakauchi S,Sumikawa K.Endogenously released ACh and exogenous nicotine differentially facilitate long-term potentiation induction in the hippocampal CA1 region of mice[J].Eur J Neurosci,2012,35(9):1381-95.
[16]Lozada AF,Wang XL,Gounko NV,et al.Induction of dendritic spines by beta 2-containing nicotinic receptors[J].J Neurosci,2012,32(24):8391-400.
[17]Bell KA,Shim H,Chen CK,et al.Nicotinic excitatory postsynaptic potentials in hippocampal CA1 interneurons are predominantly mediated by nicotinic receptors that contain alpha 4 and beta 2subunits[J].Neuropharmacology,2011,61(8):1379-88.
[18]Lombardo S,Catteau J,Besson M,et al.A role forβ2*nicotinic receptors in a model of local amyloid pathology induced in dentate gyrus[J].Neurobiol Aging,2016,46(9):221-34.
[19]Wang K,Zheng C,Wu C,et al.alpha-Chloralose diminishes gamma oscillations in rat hippocampal slices[J].Neurosci Lett,2008,441(1):66-71.
[20]Paxinos G,Franklin K.The mouse brain in stereotaxic coordinates[M].2nd ed.Elsevier:New York,2001.
[21]Yang KC,Jin GZ,Wu J.Mysterious alpha 6-containing n ACh Rs:function,pharmacology,and pathophysiology[J].Acta Pharmacol Sin,2009,30(6):740-51.
[22]Wu J,Kuo YP,George AA,et al.Beta-amyloid directly inhibits human alpha 4 beta 2-nicotinic acetylcholine receptors heterologously expressed in human SH-EP1 cells[J].J Biol Chem,2004,279(36):37842-51.
[23]Yang KC,Hu J,Lucero L,et al.Distinctive nicotinic acetylcholine receptor functional phenotypes of rat ventral tegmental area dopaminergic neurons[J].J Physiol,2009,587(2):345-61.
[24]Tozuka Y,Fukuda S,Namba T,et al.GABAergic excitation promotes neuronal differentiation in adult hippocampal progenitor cells[J].Neuron,2005,47(6):803-15.
[25]Ge S,Goh EL,Sailor KA,et al.GABA regulates synaptic integration of newly generated neurons in the adult brain[J].Nature,2006,439(7076):589-93.
[26]Cancedda L,Fiumelli H,Chen K,et al.Excitatory GABA action is essential for morphological maturation of cortical neurons in vivo[J].J Neurosci,2007,27(19):5224-35.
[27]Snyder JS,Hong NS,Mcdonald R,et al.A role for adult neurogenesis in spatial long-term memory[J].Neuroscience,2005,130(4):843-52.
[28]Winocur G,Wojtowicz JM,Sekeres M,et al.Inhibition of neurogenesis interferes with hippocampus-dependent memory function[J].Hippocampus,2006,16(3):296-304.
[29]Aimone JB,Wiles J,Gage FH.Potential role for adult neurogenesis in the encoding of time in new memories[J].Nat Neurosci,2006,9(6):723-7.
[30]Trouche S,Bontempi B,Roullet P,et al.Recruitment of adultgenerated neurons into functional hippocampal networks contributes to updating and strengthening of spatial memory[J].Proc Natl Acad Sci USA,2009,106(14):5919-24.
[31]Kitamura T,Saitoh Y,Takashima N,et al.Adult neurogenesis modulates the hippocampus-dependent period of associative fear memory[J].Cell,2009,139(4):814-27.
[32]Noonan MA,Bulin SE,Fuller DC,et al.Reduction of adult hippocampal neurogenesis confers vulnerability in an animal model of cocaine addiction[J].J Neurosci,2010,30(1):304-15.
[33]Selkoe DJ.Normal and abnormal biology of theβ-amyloid precursor protein[J].Annu Rev Neurosci,1994,17(11):489-517.
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[35]Pettit DL,Shao Z,Yakel JL.β-Amyloid1-42 peptide directly modulates nicotinic receptors in the rat hippocampal slice[J].J Neurosci,2001,21(1):1-5.
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