DISTINCT POPULATIONS OF IDENTIFIED GLIAL-CELLS IN THE DEVELOPING RAT SPINAL-CORD SLICE - ION-CHANNEL PROPERTIES AND CELL MORPHOLOGY
DISTINCT POPULATIONS OF IDENTIFIED GLIAL-CELLS IN THE DEVELOPING RAT SPINAL-CORD SLICE - ION-CHANNEL PROPERTIES AND CELL MORPHOLOGY
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DOI：
10.1111/j.1460-9568.1995.tb01027.x
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发表时间：
1995-01-01
影响因子：
3.4
通讯作者：
KETTENMANN, H
中科院分区：
文献类型：
--
作者：
CHVATAL, A;PASTOR, A;KETTENMANN, H
应用全细胞膜片钳技术，结合形态学和免疫细胞化学特征，在出生后1-19天（P1-P19）大鼠脊髓灰质中发现了四种类型的神经胶质细胞。记录的细胞用荧光黄标记，这使得随后可以使用细胞类型特异性标记物来识别细胞。胶质细胞酸性蛋白（GFAP）染色阳性。这些形态特征的多个，非常精细和短的过程和电生理对称，非衰减的K+选择性电流。少突胶质细胞被确定为一个典型的少突胶质细胞样的形态，缺乏GFAP染色和阳性标记与O 1和O 4抗体（少突胶质细胞谱系的标志物）的组合，和他们的膜是由对称的，被动的，衰减的K+电流为主。第三群胶质细胞的特征还在于O 1/O 4或仅O 4抗原阳性染色，缺乏GFAP染色，并且在一些细胞中，少突胶质细胞样形态。然而，这些细胞可以通过存在向内整流（K-IR）、延迟向外整流（K-DR）和A型K+电流（K-A）来区分，代表最可能的少突胶质细胞谱系的胶质前体细胞。第四群胶质细胞具有小胞体和广泛的长突起网络，没有明显的取向偏好。在一个案例中，过程与GFAP阳性标记，而30%的特点是微弱的，弥漫性染色。这些细胞表达复杂的电压门控通道，即Na+，K-DR，K-A和K-IR通道。与神经元相比，Na+电流的幅度至少比K+电流小一个数量级，并且这些细胞都没有表现出在电流钳模式下产生动作电位的能力。由于这些细胞都不能被少突胶质细胞标记，我们假设它们是星形胶质细胞或星形胶质细胞谱系的神经胶质前体细胞。这四种细胞类型在灰质的所有区域都有发现。当随机访问的神经胶质细胞，少突胶质细胞前体细胞和神经胶质细胞的记录与Na+电流的概率在发展过程中下降。在P1-P3，50%的细胞显示Na+电流，而在P13-P15只有18%。同时，星形胶质细胞和少突胶质细胞样膜电流的胶质细胞的数量从19和12%分别增加到41和35.5%。我们的结论是，在脊髓切片中的胶质细胞具有独特的形态，免疫组织化学和生理特性，胶质细胞群体在出生后的发展过程中发生变化。
Four types of glial cells could be distinguished in the grey matter of rat spinal cord slices at postnatal days 1-19 (P1-P19), based on their pattern of membrane currents as revealed by the whole cell patch clamp technique, and by their morphological and immunocytochemical features. The recorded cells were labelled with Lucifer Yellow, which allowed the subsequent identification of cells using cell-type-specific markers. Astrocytes were identified by positive staining for glial fibrillary acidic protein (GFAP). These were morphologically characterized by multiple, very fine and short processes and electrophysiologically by symmetrical, non-decaying K+ selective currents. Oligodendrocytes were identified by a typical oligodendrocyte-like morphology, lack of GFAP staining and positive labelling with a combination of O1 and O4 antibodies (markers of the oligodendrocyte lineage), and their membrane was dominated by symmetrical, passive, decaying K+ currents. The third population of glial cells was also characterized by positive staining for O1/O4 or only for O4 antigens, lack of GFAP staining and, in some cells, oligodendrocyte-like morphology. However, these cells could be distinguished by the presence of inwardly rectifying (K-IR), delayed outwardly rectifying (K-DR) and A-type K+ currents (K-A), representing the most likely glial precursor cells of the oligodendrocyte lineage, The fourth population of glial cells had small somata and a widespread network of long processes with no apparent orientation preference. In one case, processes were positively labelled with GFAP, while 30% were characterized by faint, diffuse staining. These cells expressed a complex pattern of voltage-gated channels, namely Na+, K-DR, K-A and K-IR channels. In contrast to neurons, the amplitude of Na+ currents was at least one order of magnitude smaller than the K+ currents, and none of these cells showed the ability to generate action potentials in the current clamp mode. Since none of these cells could be labelled by oligodendrocyte markers we assume that they were either astrocytes or glial precursor cells of the astrocyte lineage. The four cell types were found in all regions of the grey matter. When randomly accessing the glial cells, the probability of recording from the oligodendrocyte precursor cells and the glial cells with Na+ currents decreased during development. At P1-P3, 50% of the cells revealed the Na+ current, while at P13-P15 only 18% did. Concomitantly, the number of glial cells with astrocyte- and oligodendrocyte-like membrane currents increased from 19 and 12% to 41 and 35.5% respectively. We conclude that the glial cells in the spinal cord slices possess distinct morphological, immunohistochemical and physiological properties, and that the glial populations undergo changes during postnatal development.