Cholinergic neuronotrophic factors. VI. Age-dependent requirements by chick embryo ciliary ganglionic neurons.
Cholinergic neuronotrophic factors. VI. Age-dependent requirements by chick embryo ciliary ganglionic neurons.
复制标题
胆碱能神经营养因子。
DOI：
10.1016/0012-1606(81)90245-1
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发表时间：
1981
影响因子：
2.7
通讯作者：
Varon,S
中科院分区：
文献类型：
--
作者：
Manthorpe,M;Adler,R;Varon,S
在发育过程中，睫状神经节神经元成为有丝分裂后，并延长神经突起在其未来的眼内神经支配目标的存在明显独立。然而，在到达它们的外周神经支配区域后，这些神经元变得依赖于目标，其中大约一半死亡。我们以前曾报道，鸡胚眼内靶组织含有睫状神经营养因子（CNTF），它可以提取和部分纯化的可溶性形式，并确保近8天的鸡胚睫状神经节神经元在单层培养的总生存。在这项研究中，我们分离和培养睫状神经节从胚胎（艾德）5日至14日，并检查其神经元的依赖性和反应性外源性添加CNTF。两种细胞类别（暗和亮）可以区分的相位显微镜和差异计数的细胞解离ED 7 -14，但不是在ED 5 -6的。在这个发育时期，每个神经节的暗细胞数量从6000增加到78，000。相比之下，亮细胞（假定的神经元）从最大约10，000下降到6000，表明与预期的体内神经元细胞死亡相关。将来自ED 5 -14神经节的解离细胞接种在用神经突促进因子包被的聚鸟氨酸基质上，在添加或不添加CNTF的情况下培养24小时，并对存活和神经炎发展进行数值检测。ED 7 -14神经节的培养物显示出两种细胞类别：（i）扁平非神经元成分的数量随着神经节年龄的增加而急剧增加（从而与解离物中暗细胞数量的增加相关）;（ii）大而亮的细胞（通常显示出神经突生长）的数量与解离物中亮细胞数量平行减少。这些神经元元素的生存是严格依赖于外源性CNTF之间的ED 7和10，但随着年龄的增长逐渐独立。ED 14神经元（完全能够存活24小时，没有添加CNTF）继续需要CNTF的神经突延伸，从而显示保留敏感性，这一因素。虽然ED 5 -6培养物含有可识别的扁平细胞，但主要类别包括具有可变形态的细胞，几乎所有细胞都表现出神经突样突起。这些细胞的存活和轴突延伸，我们暂时解释为未成熟的神经元是独立的存在下添加的CNTF。
During development, ciliary ganglionic neurons become postmitotic and extend neurites in apparent independence of the presence of their future intraocular innervation targets. After reaching their peripheral innervation territory, however, these neurons become target dependent and about half of them die. We have previously reported that chick embryo intraocular target tissues contain a ciliary neuronotrophic factor (CNTF), which can be extracted and partially purified in a soluble form and which ensures near-total survival of 8-day chick embryo ciliary ganglionic neurons in monolayer cultures. In this study we have dissociated and cultured ciliary ganglia from embryonic Day (ED) 5 through 14, and examined dependence and responsiveness of their neurons to exogenously added CNTF. Two cell classes (dark and bright) could be distinguished by phase microscopy and differentially counted in cell dissociates from ED7–14, but not in ED5–6 ones. Dark cell number per ganglion increased from 6000 to 78,000 over this developmental time period. In contrast, bright cells (putative neurons) declined from a maximum of about 10,000 to 6000, suggesting a correlation with the expected neuronal cell deathin vivo. Dissociated cells from ED5–14 ganglia were seeded on a polyornithine substratum coated with neurite promoting factor, cultured for 24 hr with or without added CNTF, and numerically examined for survival and neuritic development. Cultures from ED7–14 ganglia showed two cell categories: (i) flat nonneuronal elements dramatically increased in number with ganglionic age (thereby correlating with the increasing number of dark cells in the dissociates) and (ii) large, bright cells (often displaying neurite outgrowth) decreased in number in parallel with bright cell number in the dissociate. The survival of these neuronal elements was strictly dependent on exogenously added CNTF between ED7 and 10, but became progressively independent with older ages. ED14 neurons (fully capable of surviving for 24 hr without added CNTF) continued to require CNTF for neurite extension, thus displaying retained sensitivity to this factor. Although the ED5–6 cultures contained well-recognizable flat cells, the dominant category comprised cells with variable morphology, practically all of which exhibited neurite-like processes. Both the survival and neurite extension of these cells, which we tentatively interpret as immature neurons were independent of the presence of added CNTF.