Culture of Hermissenda crassicornis (Mollusca) neurons.
Culture of Hermissenda crassicornis (Mollusca) neurons.
复制标题
Hermissenda crassicornis（软体动物）神经元的培养。
DOI：
10.1007/bf02634084
复制
发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Smith,PJ
中科院分区：
文献类型：
--
作者：
Tamse,CT;Collin,C;Kuzirian,AM;Smith,PJ
编辑先生：长期以来，无脊椎动物一直是中枢神经系统功能分析的宝贵模型来源。每个群体都有自己的优势，软体动物与细胞生理学和行为的研究特别相关，裸鳃类Hermissenda crassicornis在学习和记忆的研究中找到了自己的位置，特别是联想或巴甫洛夫条件反射（2）。我们对后一种系统的了解现在已经很广泛了，但由于无法培养出孤立的神经元或条件反射的回路成分而受到阻碍。Hermissenda神经元未能在培养物中生长是令人惊讶的，因为其他软体动物神经元已经成功地且相对容易地培养[例如，Aaplasia（8，11）; Helisoma（18）]，并且一些系统甚至已用于体外重建回路（12，15）。本研究描述了一种简单而可靠的技术，从Hermissenda神经元的培养，并提出了初步的数据，粘附分子的影响和电生理分离的神经元。在这项研究中特别感兴趣的细胞涉及那些在趋光回路和联合条件反射通路，例如，左足神经元（LP 1），在这个回路中特别大的运动神经元。关于这项技术的初步报告已经发表（17）。Hermissenda crassicornis是一种海洋裸鳃类软体动物，原产于美国太平洋海岸，分布范围从巴哈到阿拉斯加。它可以从Sea Life Supply，Sand City，CA获得，或者作为实验室饲养的动物从Hermissenda Resource Laboratory，Marine Biological Laboratory，伍兹霍尔，MA获得（16）。通过使用含有谷氨酰胺（ICN 12-511-54）的液体Leibovitz L-15培养基制备确定的神经元细胞培养基，并补充至终浓度/升，以mM计（括号中为g添加量）：444 NaCl（25.95 g），10.0 KCl（0.34 g），28.0 KCl（0.34 g），
Dear Editor: Invertebrates have long been an invaluable source of models for the functional analysis of the central nervous system. Each group has had its strengths, the molluscs being particularly relevant for studies of cellular physiology and behavior, with the nudibranch Hermissenda crassicornis finding its niche in the study of learning and memory, notably associative or Pavlovian conditioning (2). Our knowledge of the latter system is now extensive but has been hampered by an inability to culture the isolated neurons or circuit components of the conditioning reflex. This failure to get Hermissenda neurons to grow in culture is surprising, because other molluscan neurons have been cultured successfully and relatively easily [eg, Aplysia (8, 11); Helisoma (18)], and some systems have even been used to reconstruct circuits in vitro (12, 15). This study describes a simple and reliable technique for the culture of neurons from Hermissenda and presents preliminary data on the effect of adhesion molecules and the electrophysiology of isolated neurons. Cells of particular interest in this study involved those in the phototactic circuitry and associative conditioning pathway, for example, the left pedal neuron (LP1), a particularly large motorneuron in this circuit. A preliminary report on this technique has already been published (17). Hermissenda crassicornis, a marine nudibranch mollusc, is native to the US Pacific Coast with a range extending from Baja to Alaska. It can be obtained from Sea Life Supply, Sand City, CA, or as laboratory-reared animals from the Hermissenda Resource Laboratory, Marine Biological Laboratory, Woods Hole, MA (16). A defined neuronal cell culture medium was prepared by using liquid Leibovitz L-15 medium with glutamine (ICN 12-511-54), and supplemented to a final concentration per liter, in mM (with g ad-dition in parentheses): 444 NaCl (25.95 g), 10.0 KCl (0.34 g), 28.0