Walton M K, Schaffner A E, Barker J L
Laboratory of Neurophysiology, NIH-NINDS, Bethesda, Maryland 20892.
J Neurosci. 1993 May;13(5):2068-84. doi: 10.1523/JNEUROSCI.13-05-02068.1993.
It is not well understood when during embryonic development the elements of a cell's responsiveness first appear, nor the factors controlling their appearance. A strategy to approach this issue is to determine which aspects of neuronal development are highly stereotyped in presence, timing, or pattern across a variety of cell types, and which are more diversified by cell type, region, or other parameters. We have used a fluorescent potentiometric oxonol dye in conjunction with a digital video imaging system to record the emergence and distribution of specific forms of excitability in dissociated embryonic rat spinal cord cells. We studied the expression of responses to veratridine, a sodium channel activator; muscimol, a GABAA receptor agonist; and kainic acid, an agonist at a class of glutamate receptors. Responses were consistently detectable in a percentage of cells dissociated from the earliest age examined, embryonic day 13, and increased progressively in later ages. Cells were examined from four regions, with cervical-lumbosacral and ventrodorsal distinctions. In the population of cells from each region, functional sodium channels appeared prior to GABAA receptors, which in turn emerged prior to kainate-activated glutamate receptors. This pattern was common to all spinal cord regions and revealed ventrodorsal and rostrocaudal gradients reflecting the known pattern of spinal cord neurogenesis. Analysis of the individual cell responses indicated that the stereotypical pattern of sequential channel development occurs individually on most cells in each region.
目前尚不清楚细胞反应性的各个要素在胚胎发育的何时首次出现,也不清楚控制它们出现的因素。解决这个问题的一个策略是确定神经元发育的哪些方面在多种细胞类型中在存在、时间或模式上高度定型,哪些方面因细胞类型、区域或其他参数而更加多样化。我们使用了一种荧光电位氧杂菁染料结合数字视频成像系统,来记录解离的胚胎大鼠脊髓细胞中特定形式兴奋性的出现和分布。我们研究了对藜芦碱(一种钠通道激活剂)、蝇蕈醇(一种GABAA受体激动剂)和 kainic 酸(一类谷氨酸受体的激动剂)的反应表达。从最早检查的年龄即胚胎第13天解离的细胞中,在一定比例的细胞中始终能检测到反应,并且在随后的年龄中逐渐增加。从四个区域检查细胞,区分颈腰骶部和腹背侧。在每个区域的细胞群体中,功能性钠通道在GABAA受体之前出现,而GABAA受体又在 kainate 激活的谷氨酸受体之前出现。这种模式在所有脊髓区域都很常见,并揭示了反映已知脊髓神经发生模式的腹背侧和头尾梯度。对单个细胞反应的分析表明,通道顺序发育的定型模式在每个区域的大多数细胞上单独发生。