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胚胎期鸡交感神经元中A电流在原位及体外发育过程中的功能表达

Functional expression of A-currents in embryonic chick sympathetic neurones during development in situ and in vitro.

作者信息

Raucher S, Dryer S E

机构信息

Program in Neuroscience, Florida State University, Tallahassee, 32306-4075.

出版信息

J Physiol. 1994 Aug 15;479 ( Pt 1)(Pt 1):77-93. doi: 10.1113/jphysiol.1994.sp020279.

DOI:10.1113/jphysiol.1994.sp020279
PMID:7990036
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1155727/
Abstract
  1. The functional expression of transient voltage-activated K+ currents (IA) was examined using whole-cell recording techniques in embryonic chick sympathetic ganglion neurones that developed in situ and under various growth conditions in vitro. 2. The density of IA increased dramatically during development in sympathetic neurones isolated acutely between embryonic days 7 and 20 (E7-E20). The time course of IA inactivation became significantly faster between E7 and E13. With these protocols, neuronal differentiation and development occurred entirely in situ. 3. Sympathetic neurones isolated at E9 and maintained in vitro for 4 days did not express a normal IA compared to neurones isolated acutely at E13. Those neurones that were in physical contact with other neurones expressed normal densities of IA, but the resulting inactivation kinetics were abnormally slow. Sympathetic neurones that were cultured on the membrane fragments of lysed neurones expressed normal densities of IA even when they failed to make visible connections with other viable neurones, but the resulting inactivation kinetics were abnormally slow. Those cultured neurones that were not in physical contact with other cells or their membranes had markedly reduced densities of IA with abnormally slow inactivation kinetics. 4. Application of 5-100 ng ml-12.5 S nerve growth factor by itself did not promote normal A density of kinetics in E9 sympathetic neurones cultured for 4 days. 5. Sympathetic neurones that developed in vitro in physical contact with ventral spinal cord explants, cardiac myocytes or aortic smooth muscle cells expressed normal densities of IA, but the inactivation kinetics were abnormally slow. Cell culture media conditioned by these tissues failed to promote normal IA expression. Sympathetic neurones cultured as explants or maintained under depolarizing conditions did not express a normal IA. 6. Embryonic chick sympathetic neurones exhibit developmental changes in the density and kinetics of IA that can be regulated independently by extrinsic environmental factors including interactions with insoluble components of the plasma membranes of some cells.
摘要
  1. 运用全细胞记录技术,在原位发育以及体外不同生长条件下培养的胚胎鸡交感神经节神经元中,检测瞬时电压激活钾电流(IA)的功能表达。2. 在胚胎第7至20天(E7 - E20)急性分离的交感神经元发育过程中,IA密度显著增加。在E7至E13期间,IA失活的时间进程明显加快。通过这些实验方案,神经元分化和发育完全在原位发生。3. 与E13急性分离的神经元相比,E9分离并在体外培养4天的交感神经元未表达正常的IA。那些与其他神经元有物理接触的神经元表达正常密度的IA,但由此产生的失活动力学异常缓慢。在裂解神经元的膜片段上培养的交感神经元,即使它们未能与其他活神经元建立可见连接,也表达正常密度的IA,但由此产生的失活动力学异常缓慢。那些未与其他细胞或其膜有物理接触的培养神经元,IA密度显著降低,失活动力学异常缓慢。4. 单独应用5 - 100 ng/ml的2.5S神经生长因子,并未促进培养4天的E9交感神经元中正常的IA密度动力学。5. 在体外与腹侧脊髓外植体、心肌细胞或主动脉平滑肌细胞有物理接触而发育的交感神经元,表达正常密度的IA,但失活动力学异常缓慢。这些组织条件培养液未能促进正常的IA表达。作为外植体培养或在去极化条件下维持的交感神经元未表达正常的IA。6. 胚胎鸡交感神经元在IA密度和动力学方面表现出发育变化,这些变化可由外在环境因素独立调节,包括与某些细胞质膜不溶性成分的相互作用。

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