创伤后应激障碍对小鼠背侧及腹侧海马谷氨酸能和GABA能神经元兴奋性的影响

本研究旨在通过检测创伤后应激障碍(post-traumatic stress disorder, PTSD)小鼠背侧海马(dorsal hippocampus, dHPC)及腹侧海马(ventral hippocampus, vHPC)神经元电生理特性的改变,探讨海马神经元的可塑性与PTSD后记忆之间的关系。随机将C57Thy1-YFP/GAD67-GFP品系雄性小鼠分为对照组和PTSD组,通过对小鼠施加不可逃避足底电击(foot shock, FS)建立PTSD模型,利用Morris水迷宫探查小鼠空间学习记忆的变化,在全细胞记录模式下检测dHPC和vHPC谷氨酸能神经元和GABA能神经元电生理特性的变化。结果显示：FS可显著降低小鼠平均移动速度、提高僵立次数和僵立百分比;PTSD可显著延长定位逃避训练中的定位逃避期,缩短定位探查训练中在原象限内游泳停留时间,延长在对侧象限内游泳停留时间;FS同时提高dHPC谷氨酸能神经元和vHPC GABA能神经元能障、绝对不应期和动作电位间距,降低dHPC GABA能神经元和vHPC谷氨酸能神经元能障、绝对不应期和动作电位间距。以上结果表明：PTS...     

BDNF及其下游通路与GABA能神经元发育相关性的研究进展

脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)是一种具有神经营养作用的蛋白质,广泛分布于中枢神经系统内。BDNF及其下游信号通路在γ-氨基丁酸(γ-aminobutyric acid, GABA)能神经元存活、生长、分化、发育等方面均发挥重要的作用。GABA能神经元可以通过释放抑制性神经递质GABA调节神经元活性,进而维持神经环路的正常功能。多种疾病的发生发展都与GABA能神经元发育的异常密切相关。该文将就BDNF及其下游通路与GABA能神经元发育的相关性进行综述,希望为疾病的治疗提供新的方向。     

猕猴颈髓的GABA能神经元分布

应用免疫组织化学方法观察猕猴颈髓的γ-氨基丁酸（GABA）能神经元的分布,观察结果：除第X层外,在脊髓RexedⅠ-Ⅸ层可见GABA样免疫反应的胞体和纤维,标记的GABA胞体为卵圆形,三角形和多角形,可分为大、中、小型,在Ⅲ、Ⅳ、Ⅴ、Ⅶ、Ⅸ层GABA阳性胞体较多,GABA阳性纤维以后角处最多,白质内也有GABA免疫反应阳性的胶质细胞和神经纤维。结果提示GABA能神经元不仅调节感觉信息的传导而且也调节运动信息的传导。     

猫下丘中央核GABA能神经元年龄相关变化

目的比较研究猫下丘中央核(CIC)GABA能神经元年龄相关变化,探索老年个体听力下降的神经机制。方法Nissl染色显示下丘神经元,免疫组织化学ABC法显示γ-氨基丁酸(GABA)免疫阳性神经元。光镜下观察、拍照,对神经元和GABA能神经元分别计数并换算成密度,测量GABA能神经元直径取平均值。结果GABA阳性反应神经元、阳性纤维及其终末在青年猫及老年猫下丘中央核均有分布。与青年猫相比,老年猫下丘中央核神经元及GABA能神经元密度均显著下降(P<0.01),GABA能神经元下降幅度较大;GABA能神经元胞体直径明显减小(P<0.01),阳性反应明显减弱。结论在衰老过程中猫下丘神经元尤其是GABA能神经元有显著丢失现象,提示GABA能神经元显著减少导致下丘兴奋性和抑制性神经递质之间的平衡失调,可能是引起老年个体听觉功能衰退的重要原因。     

猫上丘浅层GABA能神经元的年龄相关性变化

目的比较青年猫和老年猫上丘浅层（superricial Superior Colliculus,sSC）GABA能神经元及其表达的年龄相关性变化,探讨老年个体视觉功能衰退的相关神经机理。方法Nissl染色显示上丘浅层结构及神经元、免疫组织化学ABC法标记GABA免疫阳性神经元。光镜下观察,采集图像,并利用图像分析软件对带状层、浅灰质层和视层神经元及GABA免疫阳性神经元及其灰度值进行分析统计。结果GABA免疫阳性神经元、阳性纤维及其终末在青年猫及老年猫上丘浅层均有分布。与青年猫相比,老年猫上丘浅灰质层、视层神经元和GABA免疫阳性神经元密度及其GABA免疫阳性反应强度均显著下降（P〈0．01）（免疫反应强度与平均灰度值成反比）;带状层神经元密度也显著下降（P〈0．01）,但其GABA免疫阳性神经元密度无显著变化（P〉0．05）。结论衰老过程中猫上丘浅层GABA能神经元的丢失和GABA表达的下降,可能是在上丘水平上导致老年个体视觉功能衰退的重要因素之一。     

猫腰髓中GABA能神经元的分布及年龄相关变化

目的观察和比较GABA能神经元在青年猫和老年猫L6段脊髓的分布,探讨GABA能神经元在脊髓中分布的年龄相关变化及意义.方法免疫组织化学ABC法.结果青年猫与老年猫L6段脊髓灰质内,GABA能神经元及神经纤维分布广泛,各个Rexed板层均可见GABA-IR细胞,其中背侧灰质阳性最强,其次是腹侧灰质.标记的GABA能神经元胞体为卵圆形、三角形、多角形和星形,可分为大、中、小三种类型.经比较,老年组GABA能神经元的数量及免疫反应性均明显低于青年组.结论老年动物脊髓调节功能的减弱可能与GABA能神经元减少有少.     

KF核及B(o)tzinger复合体内GABA能神经元向膈神经核的投射

实验在６只成年猫上进行。将ＷＧＡ－ＨＲＰ微量注入Ｃ５膈神经核内,通过逆行追踪及ＧＡＢＡ免疫组织化学ＦＩＴＣ荧光双重标记方法,研究了脑干内ＧＡＢＡ能神经元向膈神经核的投射。结果在脑桥ＫＦ核和面神经后核周围区（即Ｂｏｔｚｉｎｇｅｒ复合体）观察到ＧＡＢＡ－ＨＲＰ双标神经元。另外,在中缝大核、旁巨细胞外侧核及前庭神经核也观察到双标神经元。本实验结果表明：发自上述脑干神经核团,特别是ＫＦ核及Ｂｏｔｚｉｎｇｅ     

猫小脑皮质GABA能神经元年龄相关性变化

为探讨青年猫和老年猫小脑皮质GABA能神经元及其表达的年龄相关性变化,利用Nissl染色显示小脑皮质结构及神经元,免疫组织化学ABC法标记GABA免疫阳性神经元。光镜下观察,采集图像,并利用图像分析软件对分子层、蒲肯野细胞层和颗粒层神经元及GABA免疫阳性神经元及其灰度值进行分析统计。结果显示,GABA免疫阳性神经元、阳性纤维及终末在青年猫和老年猫小脑皮质各层均有分布。与青年猫相比,老年猫分子层、蒲肯野细胞层神经元和GABA免疫阳性神经元密度及其GABA免疫阳性反应强度均显著下降(P<0.01),颗粒层神经元密度和GABA免疫阳性强度也显著下降(P<0.01),但其GABA免疫阳性神经元密度无显著变化(P>0.05);蒲肯野细胞的胞体萎缩,阳性树突分枝减少。因此认为,衰老过程中猫小脑皮质GABA能神经元的丢失和GABA表达的下降,可能是老年个体运动协调、精确调速和运动学习等能力下降的重要原因之一。     

螽斯听觉中间神经元TN2的声反应特征以及GABA对其影响

采用单细胞电生理记录技术,对螽斯Ｇａｍｐｏｃｌｅｉｓ ｇｒａｔｉｏｓａ听觉双轴突中间神经元ＴＮ２的声反应放电活动的基本特征进行了观测,发现ＴＮ２的放电模式为“ｐｈａｓｉｃ”型,最敏感频率为１３ｋＨｚ,反应阈值为３１ｄＢ ＳＰＬ,是一个高灵敏、宽带通的神经元。还研究了抑制性神经递质ＧＡＢＡ及其拮抗剂苦毒素对ＴＮ２声反应的影响,发现ＧＡＢＡ能抑制ＴＮ２的放电活动,而苦毒素则将其放电模式改变为“ｔｏｎｉ     

KF核及Btzinger复合体内GABA能神经元向膈神经核的投射

实验在 6只成年猫上进行。将WGA HRP微量注入C5膈神经核内 ,通过逆行追踪及GABA免疫组织化学FITC荧光双重标记方法 ,研究了脑干内GABA能神经元向膈神经核的投射。结果在脑桥KF核和面神经后核周围区 (即B¨otzinger复合体 )观察到GABA HRP双标神经元。另外 ,在中缝大核、旁巨细胞外侧核及前庭神经核也观察到双标神经元。本实验结果表明 :发自上述脑干神经核团 ,特别是KF核及B¨otzinger复合体的GABA能神经元的轴突可投射到膈神经核     

The postnatal development of intrinsic properties and spike encoding at cortical GABAergic neurons

GABAergic neurons play a critical role in maintaining the homeostasis of brain functions for well-organized behaviors. It is not known about the dynamical change in signal encoding at these neurons during postnatal development. We investigated this issue at GFP-labeled GABAergic neurons by whole-cell recording in cortical slices of mice. Our results show that the ability of spike encoding at GABAergic neurons is improved during postnatal development. This change is associated with the reduction of refractory periods and threshold potentials of sequential spikes, as well as the improvement of linear correlations between intrinsic properties and spike capacity. Therefore, the postnatal maturation of the spike encoding capacity at GABAergic neurons will stabilize the excitatory state of cerebral cortex.     

Ontogeny of Neurochemical Markers for Noradrenergic, GABAergic and Cholinergic Neurons in Neocortex Lesioned with Methylazoxymethanol Acetate

Abstract: Methylazoxymethanol acetate (MAM), a potent, rapidly eliminated nucleic acid alkylating agent, produces microencephaly in rat pups when injected into their dams on day 15 of gestation. In the adult microencephalic rats, neuronal loss is largely confined to telencephalic structures, such as the superficial neocortical laminae, whose neuroepithelial progenitor cells were undergoing vigorous replication during the chemical exposure. Histological examination of the forebrain 2 days after injection revealed early selective damage to the ventricular geminal zone with relative sparing of cortical plate neurons generated on earlier days. The degree of specificity of MAM's action on neurochemically defined neuronal populations was examined by measuring presynaptic markers for GABAergic, noradrenergic and cholinergic neurons in atrophic lateral cortex from 20 days gestation to adulthood. Although treatment reduced GABAergic markers (GABA, its synthetic enzyme and synaptosomal uptake process) in proportion to loss of cortex mass (-67%), the maturational pattern for remaining GABAergic neurons was virtually normal. Although the maturational sequence of noradrenergic markers was similar to control, the concentration of endogenous norepinephrine, [³H]norepinephrine uptake and tyrosine hydroxylase specific activity were two- to fourfold higher than control at each time. However, total noradrenergic markers per cortex section were nearly identical to control throughout development, indicating that development of the noradrenergic axonal arbor in neocortex was insensitive to loss of neurons in the terminal field. Maturation of cholinergic markers (endogenous acetylcholine, its synthetic enzyme and [³H]choline uptake) in the atrophic cortex was biphasic: concentrations were similar to control values for the first 12 postnatal days, but gradually rose to levels twofold higher than control. These results indicate that neurochemical alterations observed in cortex from prenatally MAM-treated rats are primarily the result of early selective elimination of neuronal subpopulations. Fetal MAM exposure appeared to have minimal effects on biochemical differentiation of neurons remaining intact in the atrophic cortex. MAM appears to be a useful toxin for producing selective loss of neuronal groups based on their time of generation in the fetus.     

Intracellular Ca regulates spike encoding at cortical GABAergic neurons and cerebellar Purkinje cells differently

Spike encoding at GABAergic neurons plays an important role in maintaining the homeostasis of brain functions for well-organized behaviors. The rise of intracellular Ca²⁺ in GABAergic neurons causes synaptic plasticity. It is not clear how intracellular Ca²⁺ influences their spike encoding. We have investigated this issue at GFP-labeled GABAergic cortical neurons and cerebellar Purkinje cells by whole-cell recording in mouse brain slices. Our results show that an elevation of intracellular Ca²⁺ by infusing adenophostin-A lowers spike encoding at GABAergic cortical neurons and enhances encoding ability at cerebellar Purkinje cells. These differential effects of cytoplasmic Ca²⁺ on spike encoding are mechanistically associated with Ca²⁺-induced changes in the refractory periods and threshold potentials of sequential spikes, as well as with various expression ratios of CaM-KII to calcineurin in GABAergic cortical neurons and cerebellar Purkinje cells.     

Neurochemistry of GABAergic System in Cerebral Cortex Chronically Exposed to Bromide In Vivo

Neurochemical correlates of GABAergic synaptic transmission [binding, uptake, metabolism, and tissue content of gamma-aminobutyric acid (GABA)] were investigated in the cortex of rats that had been given 27 mM bromide in drinking water for periods of time ranging from 1 day to 1 month. No effect of bromide on any of the parameters was found and it is concluded that chronic administration of bromide has no profound effect on GABAergic inhibitory system in the rat cortex.     

Function of GABAergic and glutamatergic neurons in the stomach

-Aminobutyric acid (GABA) and L-glutamic acid (L-Glu) are transmitters of GABAergic and glutamatergic neurons in the enteric interneurons, targeting excitatory or inhibitory GABA receptors or glutamate receptors that modulate gastric motility and mucosal function. GABAergic and glutamatergic neuron immunoreactivity have been found in cholinergic enteric neurons in the stomach. GABA and L-Glu may also subserve hormonal and paracrine signaling. Disruption in gastrointestinal function following perturbation of enteric GABA receptors and glutamate receptors presents potential new target sites for drug development.     

Bone Morphogenetic Protein-2 Promotes Survival and Differentiation of Striatal GABAergic Neurons in the Absence of Glial Cell Proliferation

We examined the potential neurotrophic effects of bone morphogenetic protein (BMP)-2 on the survival and differentiation of neurons cultured from the rat developing striatum at embryonic day 16, a period during which the mRNAs for BMP-2 and its receptor subunits (types IA, IB, and II) were detected. BMP-2 exerted potent activity to promote the survival of striatal neurons and increased the number of surviving microtubule-associated protein-2-positive cells by 2.4-fold as compared with the control cultures after 4 days in vitro. Although basic fibroblast growth factor (bFGF) also showed relatively high activity to promote the survival of striatal neurons, transforming growth factor-beta1, -beta2, and -beta3, glial cell line-derived neurotrophic factor, or brain-derived neurotrophic factor promoted their survival weakly. Striatal neurons cultured in the presence of BMP-2 or bFGF possessed extensive neurite outgrowths, the majority of which were GABA-immunoreactive. Inhibition of glial cell proliferation by 5-fluorodeoxyuridine did not affect the capacity of BMP-2 to promote the survival of striatal GABAergic neurons. In contrast, the ability of bFGF to promote the survival of striatal neurons was inhibited significantly by the treatment of cells with 5-fluorodeoxyuridine. All these results suggest that BMP-2 exerts potent neurotrophic effects on the striatal GABAergic neurons in a glial cell-independent manner.     

Differential sensitivity of cholinergic and GABAergic neurons in chick embryos treated intracerebrally with ethanol at 8 days of embryonic age

We have shown that in embryos treated with ethanol in ovo during days 1–3, a critical period of neuroembryogenesis, cholinergic neuronal phenotypic expression is decreased whereas GABAergic and catecholaminergic neuronal populations are increased as assessed by neuronal markers choline acetyltransferse (ChAT), glutamic acid decarboxylase (GAD) and tyrosine hydroxylase (TH) respectively. In this study, ethanol was administered intracerebrally to embryos at embryonic day 8, embryos were sacrificed at day 9 and ChAT and GAD activities assayed separately in cerebral hemispheres and remaining brain (diencephalon-midbrain and optic lobes). We found that ChAT activity was enhanced in the cerebral hemispheres only, whereas GAD activity was decreased in both cerebral hemispheres and remaining brain. We have concluded that the differential responses of neuronal phenotypes to ethanol may reflect compensatory mechanisms to ethanol insult. Moreover, these findings emphasize the vulnerability of the GABAergic neuronal phenotypes to ethanol neurotoxicity during early brain development in the chick.     

大脑皮层神经元膜蛋白构象改变的ESR研究

采用马来酰亚胺标记完整的大脑皮层细胞,观察由低氧引起的ESR谱线的变化及脑细胞脂质过氧化程度,低氧引起细胞过氧化物生成增加,膜蛋白构象改变．金属硫蛋白(10^-5mol/L)能明显抑制过氧化反应,具有一定的抗氧化作用．并对实验的分子机理进行了简要的讨论．     

山羊胚胎大脑皮层神经干细胞分离、培养与鉴定

目的 :从山羊胚胎大脑皮层中分离培养并鉴定神经干细胞。方法 :利用NBS培养和单细胞克隆技术在山羊胚胎大脑皮层中分离出具有单细胞克隆能力的细胞 ,并进行培养、传代、分化观察 ,采用免疫组化检测克隆细胞的神经巢蛋白 (Nestin)抗原和分化后特异性成熟神经细胞抗原的表达。结果 :从胚龄 2 4～ 30d的新鲜山羊胚胎大脑皮层中成功分离出神经干细胞 ,该细胞具有连续克隆能力 ,可传代培养 ,表达神经巢蛋白抗原。分化后的细胞表达神经元细胞、胶质细胞和少突胶质细胞的特异性抗原。结论 :山羊胚胎大脑皮层中存在具有自我更新能力和多分化潜能的神经干细胞。     

皮层SI区伤害感受神经元膜电生理特性研究

用细胞内记录技术, 在16只成年健康猫, 研究了皮层第一躯体感觉区(primary somatosensory cortex area,SI区)伤害感受神经元的电生理特性.SI区伤害感受神经元自发放电频率差异大,放电形式多样.极化电流绝对值≤1.0 nA时,伤害感受神经元I-V极相关(r=0.96),整流作用不明显;极化电流绝对值＞1.0 nA时,在两个方向上发生整流,I-V(电流-电压)曲线表现为“S”型, 其中伤害感受神经元整流作用较非伤害感受神经元明显.伤害感受神经元R_m、τ、C_m明显大于非伤害感受神经元(P＜0.01或P＜0.05).结果提示SI区伤害感受神经元与非伤害感受神经元可能在细胞膜结构、细胞大小等方面存在有意义的差别,从而反映其不同的生理功能.此电学参数特点也可为痛觉的特异性学说提供实验资料.