Kirchhof B, Bicker G
Institut für Neurobiologie, Freie Universität Berlin, Federal Republic of Germany.
J Comp Neurol. 1992 Sep 15;323(3):411-22. doi: 10.1002/cne.903230308.
We developed a cell culture system for thoracic neurons of fifth instar or adult locusts (Locusta migratoria) in order to obtain maximum visualization of cellular morphology and direct access to the neurons for electrophysiological analysis. The dissociated neurons regenerated new neurites in a serum-free defined culture medium, in which they remained viable for up to 3 weeks. Viability of the cells was confirmed by intracellular recordings demonstrating active membrane properties and action potentials. While the morphology of the cultured neurons is distinct from their in vivo counterparts, they retained some cellular surface properties and markers related to transmitter metabolism. Two factors influencing cellular morphology in vitro were identified in Locusta: 1) the presence of a primary neurite stump, and 2) membrane contacts between cells. Dissociated neurons of the locust species Schistocerca gregaria grown in a hemolymph-enriched medium showed a marked reduction in branching patterns and a tenfold increase in neurite length compared to neurons growing in a medium without hemolymph. This culture system could prove useful for identifying the action of hemolymph-derived growth factors.
我们开发了一种用于五龄或成年蝗虫（飞蝗）胸神经元的细胞培养系统，以便最大程度地观察细胞形态，并直接对神经元进行电生理分析。解离后的神经元在无血清的限定培养基中再生出新的神经突，它们在这种培养基中可存活长达3周。通过细胞内记录证实细胞具有活性膜特性和动作电位，从而确认了细胞的活力。虽然培养神经元的形态与其体内对应物不同，但它们保留了一些与递质代谢相关的细胞表面特性和标记物。在飞蝗中确定了影响体外细胞形态的两个因素：1）初级神经突残端的存在，以及2）细胞之间的膜接触。与在无血淋巴培养基中生长的神经元相比，在富含血淋巴的培养基中生长的沙漠蝗解离神经元的分支模式明显减少，神经突长度增加了十倍。这种培养系统可能有助于确定血淋巴衍生生长因子的作用。
