红螯螯虾XO-SG神经细胞的原代培养  

Primary cell culture of XO-SG from red claw crayfish(Cherax quadricarinatus)

作　　者：张岩[1] 段虎[1] 金松君[1] 李富花[1] 相建海[1] 

机构地区：[1]中国科学院海洋研究所,实验海洋生物学重点实验室,山东青岛266071

出　　处：《水产学报》2016年第10期1606-1612,共7页Journal of Fisheries of China

基　　金：国家重点基础研究发展计划(2012CB114403);国家基金国际合作交流项目(31461143007);国家自然科学基金(41376165)

摘　　要：本研究采用酶解的方法获得红螯螯虾眼柄XO-SG复合体的单个神经细胞,并依据显微观察对XO-SG神经细胞进行分类,同时利用Leibovitz's L-15等作为基础培养基离体培养解离的神经细胞。目的是建立虾类XO-SG神经细胞的分类标准并确定合适的培养条件,便于在体内外开展神经内分泌系统的调控研究。结果显示,根据神经细胞形态特点,红螯螯虾XO-SG神经内分泌细胞分为6种类型,不同类型的细胞在细胞大小以及显微结构上存在明显差异;建立了红螯螯虾眼柄XO-SG神经元的体外原代培养方法,细胞在改良的L-15培养基中存活状态良好,原代培养的神经细胞可以在体外存活14 d。离体培养过程中,不同类型神经细胞的再生速率存在差异,部分细胞在第2天就出现再生的轴突或树突。再生过程基本可以持续7 d,随后细胞开始萎缩凋亡。本研究为红螯螯虾眼柄神经细胞的分类以及利用神经细胞在体外开展虾类的内分泌代谢和调节机制研究提供了基础依据和研究平台。In this study, eyestalks of red claw crayfish were dissociated with trypsin to gain single neurons and classified neurons by microscopic inspection. And then, dissociated neurons were cultured into sterile Leibovitz＇s L-15 medium to screen out proper conditions, etc. The goal of the study is to establish a standard for neurons classification and carry out further research about the endocrine system regulation for crustacean in vitro and in vivo. The results showed that eyestalk XO-SG neurons of red claw crayfish, Cherax quadricarinatus were classified into six different types based on morphological characters with differences on the cell size, and some other differences at microscopic levels. A cell culture approach has been established for XO-SG neurons in vitro,and neurons showed an immediate outgrowth and could regenerate in modified L-15 medium for axon or dendrite.Primary neurons could survive more than 14 d with different growth rate. Some neurons exhibit regeneration of axon or dendrite two days later. Regenerative process can last for 7 days, from then on, the neurons start to shrink and apoptosis. This research provided basis for further study in classifying XO-SG neurons and understanding the endocrine metabolism and regulation at the cell level.

关 键 词：红螯螯虾 眼柄 原代培养 神经细胞 再生 

分 类 号：S917.4[农业科学—水产科学]