Guthrie P B, Brenneman D E, Neale E A
Laboratory of Developmental Neurobiology, National Institute of Child Health and Human Development, Bethesda, MD 20892.
Brain Res. 1987 Sep 15;420(2):313-23. doi: 10.1016/0006-8993(87)91252-2.
The neuronal properties of separate dissociated cell cultures of dorsal and ventral halves of the embryonic mouse spinal cord (E 13.5) were investigated. Ventral-half cultures grew on a variety of substrates and in a variety of media; dorsal-half cultures required a non-neuronal feeder layer and supplemented medium for survival. The two types of cultures differed in their morphological and biochemical properties. Ventral-half neurons remained well separated on the culture plate, whereas dorsal-half neurons tended to aggregate. Lucifer yellow fills showed that ventral-half neurons were substantially larger and had more processes than dorsal-half neurons. Because of the large size and good separation of the neurons, ventral-half cultures provide an especially attractive system for electrophysiologic and morphologic studies. Ventral-half cultures were highly enriched for choline acetyltransferase (ChAT) activity and had more neurons that stained for intracellular acetylcholinesterase (AChE); dorsal-half cultures were enriched for glutamic acid decarboxylase (GAD) activity, and high-affinity gamma-aminobutyric acid (GABA) uptake. The clear differences between the two cultures indicate that many morphological and biochemical properties are already specified on embryonic day 13.5.
对胚胎小鼠脊髓(E13.5)背侧和腹侧分离的解离细胞培养物的神经元特性进行了研究。腹侧半培养物在多种底物和多种培养基上生长;背侧半培养物需要非神经元饲养层和补充培养基才能存活。这两种培养物在形态和生化特性上有所不同。腹侧半神经元在培养板上保持良好的分离状态,而背侧半神经元则倾向于聚集。荧光黄填充显示,腹侧半神经元比背侧半神经元大得多,且有更多的突起。由于神经元体积大且分离良好,腹侧半培养物为电生理和形态学研究提供了一个特别有吸引力的系统。腹侧半培养物中胆碱乙酰转移酶(ChAT)活性高度富集,且有更多神经元对细胞内乙酰胆碱酯酶(AChE)染色;背侧半培养物中谷氨酸脱羧酶(GAD)活性富集,且有高亲和力的γ-氨基丁酸(GABA)摄取。两种培养物之间的明显差异表明,许多形态和生化特性在胚胎第13.5天就已经确定。