摘要
香鱼(Plecoglossus altivelis)是典型的降海洄游鱼类,由于地理原因,一些香鱼在繁殖季无法回到海中产卵,形成陆封型香鱼。陆封型和洄游型香鱼相比,其体型明显偏小。鱼类的体肾组织可以调控渗透压,从而影响生长,因此我们采用蛋白质组学的方法研究陆封型和洄游型香鱼体肾组织蛋白质组的差异。双向电泳(2-DE)分析筛选到25个差异表达蛋白点,质谱分析成功鉴定出21个蛋白。与洄游型香鱼相比,陆封型香鱼体肾组织中NADH脱氢酶、异柠檬酸脱氢酶、α-微管蛋白、热适应相关蛋白65(Wap65)、热休克蛋白60(HSP60)、丙酮酸脱氢酶(E1)、碳酸酐酶、烯醇酶和乳酸脱氢酶表达量较高。同时陆封型香鱼的β-肌动蛋白、铁蛋白、氨基酰化酶、甲硫氨酸腺苷转移酶、过氧化物酶、丙氨酸乙醛酸转移酶和磷酸烯醇丙酮酸羧激酶则表达量较低。它们主要参与能量代谢、应激反应、氨基酸代谢等过程。由于Wap65、HSP60和E1 3种蛋白在蛋白质组学结果中表达改变,且与应激和能量代谢有关,可能影响鱼类生长,因此采用荧光定量PCR(RT-PCR)验证陆封型和洄游型香鱼体肾Wap65、HSP60和E1基因mRNA表达差异,结果表明这3个基因mRNA表达均在陆封型香鱼体肾中较高,与蛋白质组学结果一致。综上,陆封型香鱼的几个糖酵解相关酶和应激蛋白表达量比洄游型香鱼高,而蛋白代谢相关酶表达量较低,揭示陆封型和洄游型香鱼在环境应激和能量代谢等层面有明显的差异。
Ayu （Plecoglossus altivelis） is a typical amphidromous fish. Since some ayu can not return to sea to spawn during breeding season, they became land-locked type ayu. The body size of land-locked type ayu is smaller compared with that of amphidromous type ayu. Trunk kidney is an essential organ for fish osmoregulation, it may influence fish growth. The differences of trunk kidney proteome between land-locked type and amphidromous type ayu were investigated. Twenty-five protein dots were found using two dimension elec- trophoresis （2-DE）, and twenty-one dots of them were successfully identified by mass spectrum. Compared to those in amphidromous type ayu, the expression of NADH dehydrogenase, isocitrate dehydrogenase, o^-tubulin, heat shock 60 kDa protein （ HSP60）, pyru- vate dehydrogenase （E1）, carbonic anhydrase, warm temperature acclimation related 65 kDa protein （Wap65）, enolase and lactate dehydrogenaseareinare were higher in land-locked type ayu, whereas alanine-glyoxylate aminotransferase, 13-actin, aminoacylase, phos- phoenolpyruvate carboxykinase, ferritin H3, methionine adenosyhransferase and peroxiredoxin were lower in land-locked type ayu. These proteins were involved mainly in the processes of energy metabolism, responses to stress and amino acid metabolism. Wap65, HSP60, and E1 were changed in protein expression levels related with stress and energy metabolism. Real-time PCR （RT-PCR） was employed to measure the expression of Wap65, HSP60 and E1 genes between land-locked type and amphidromous type ayu. The data showed that mRNA expression of three selected genes were relatively high in land-locked type ayu.
出处
《生物学杂志》
CAS
CSCD
2014年第2期13-17,共5页
Journal of Biology
基金
国家自然科学基金项目(31201970)
关键词
香鱼
洄游型
陆封型
蛋白质组学
差异表达
ayu
amphidromous type
land-locked type
proteome analysis
differential expression
