CHARACTERIZATION OF GABAERGIC NEURONS IN HIPPOCAMPAL CELL-CULTURES
CHARACTERIZATION OF GABAERGIC NEURONS IN HIPPOCAMPAL CELL-CULTURES
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DOI：
10.1007/bf01188497
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发表时间：
1994-05-01
期刊：
影响因子：
--
通讯作者：
BANKER, G
中科院分区：
文献类型：
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作者：
BENSON, DL;WATKINS, FH;BANKER, G
体外培养4~35d的海马神经元，观察GABA能神经元的形态特征和GABA能突触终末的分布。在所有时间点，表达GABA免疫反应的神经元约占培养神经元总数的6%。尽管GABA能细胞的形态特征表明是一个异质性的群体，但GABA能细胞作为一类明显不同于非GABA能细胞，可能是锥体细胞。大多数GABA能细胞比非GABA能细胞具有更多的梭形或多边形的胞体，无刺和较少的锥形树突，并且表现出更高的相密度。定量分析表明，与非GABA能神经元相比，GABA能细胞具有更少的初级树突、更细长的树突和更长的树突节段，这些特征与原位GABA能细胞相似。双重免疫染色显示，GAD65阳性的精索静脉曲张对突触蛋白I也呈免疫阳性，提示GAD65阳性的精索静脉曲张接触胞体和树突代表突触前特化。共聚焦显微镜显示，胞体上GAD65阳性的突触专化的比例大于树突，这表明体细胞和树突诱导GABA能轴突形成突触前专化的能力不同。这些数据表明，在培养中发育的GABA能细胞表现出与非GABA细胞不同的形态特征，并参与不同的突触相互作用。因此，在培养中区分GABA能神经元的许多特征使人想起区别原位GABA能神经元的特征。
The morphological characteristics of GABAergic neurons and the distribution of GABAergic synaptic terminals were examined in cultures of hippocampal neurons from 4-35 days in vitro. Neurons expressing GABA immunoreactivity represented about 6% of the total number of cultured neurons at all time points. Although the morphological characteristics of GABAergic cells suggested a heterogeneous population, GABAergic cells as a class were notably different from the non-GABAergic, presumably pyramidal cells. Most GABAergic cells had more fusiform or polygonal shaped somata, non-spiny and less tapering dendrites and appeared more phase-dense than nonGABAergic cells. Quantitative analysis revealed that GABAergic cells had fewer primary dendrites, more elongated dendritic arbors, and longer dendritic segments than non-GABAergic neurons-characteristics that are similar to GABAergic cells in situ. Double immunostaining revealed that GAD65-positive varicosities were also immunopositive for synapsin I, suggesting that GAD65-positive varicosities that contacted somata and dendrites represented presynaptic specializations. Confocal microscopy revealed the proportion of the synaptic specializations on the cell soma that were GAD65-positive was greater than on the dendrites, suggesting that somata and dendrites differ in their ability to induce the formation of presynaptic specializations by GABAergic axons. These data indicate that the GABAergic cells that develop in culture exhibit distinctive morphological characteristics and participate in different synaptic interactions than nonGABA cells. Thus many of the features that distinguish GABAergic neurons in culture are reminiscent of the characteristics that distinguish GABAergic neurons in situ.