高钙和缺钙环境对花鳗鲡血液生理指标及其NCXs和PMCAs基因表达的影响  

Effects of high calcium and calcium deficiency on blood physiological indexes and mRNA expression of NCXs and PMCAs in Anguilla marmorata

作　　者：曹全全 胡亚东 王涛 张宏叶 顾杰 王丹[1] 李欣茹 尹绍武[1] 

机构地区：[1]南京师范大学生命科学学院海洋科学与工程学院 [2]江苏大学生命科学学院,江苏南京210023 [2]江苏大学生命科学学院,江苏镇江212000

出　　处：《海洋渔业》2018年第4期424-433,共10页Marine Fisheries

基　　金：江苏省自然科学基金项目(BK20141450)

摘　　要：为探究花鳗鲡(Anguilla marmorata)应对极性钙浓度环境下体内的应激机制,检测了花鳗鲡成鳗从对照组(2 mM Ca^(2+))转移到高钙组(10 mM Ca^(2+))和缺钙组(0 mM Ca^(2+))时第1、4和7天的血清钙浓度、血清渗透压和5项血液指标(白细胞数目、红细胞数目、葡萄糖浓度、血红蛋白浓度和血小板数目),采用qRT-PCR技术对NCXs和PMCAs基因在花鳗鲡不同组织中的表达进行分析,并研究了花鳗鲡应对极性钙浓度下NCXs和PMCAs基因在鳃组织中表达变化规律。结果表明:血清钙浓度和血清渗透压与周围钙离子浓度成正相关,第1、4天处理组较对照组有显著性差异(P<0.05),第7天不同组之间无显著差异(P>0.05);随时间延长高钙环境下各项指标由高到低,低钙环境下各项指标由低到高,至第7天达到稳态;5项血液指标,第1、4天各处理组数值均比对照组高,第7天达到稳态。对于组织表达,NCX2和NCX3在皮肤中表达最高,而NCX1、PMCA2和PMCA4在肾脏中表达最高。对于各亚型的mRNA时序表达,NCX1和PMCA4在极性钙处理情况下相较对照组有显著性差异(P<0.05);其中,NCX1在高钙条件下随着时间增长表达量逐渐升高,在低钙环境下随着时间增长表达量逐渐降低,即NCX1的表达对于极性钙环境具有指示作用;PMCA4在高钙条件下随着时间延长表达量逐渐降低,在低钙环境下随着时间延长表达量逐渐升高,在第7天达到稳态,即PMCA4的表达对于钙环境的稳态具有指示作用;其它亚型随着时间的延长均无显著性差异。可见NCX1和PMCA4在花鳗鲡洄游过程机体内钙稳态的调节中扮演了重要角色。Calcium plays an important part in sea water,and its concentration fluctuates widely in the surface and deep water of the sea. But the calcium concentrations in fish need to be kept within a narrow range. As an important catadromous fish, marbled eels（ Anguilla marmorata） need to adapt to different Ca^（2＋）concentrations. In order to explore the stress mechanism of eels in the acute calcium concentration water environment,we measured the serum calcium concentration,serum osmotic pressure and five blood indexes from the control,high calcium and calcium deficiency group. Meanwhile,we explored the tissue distribution of NCXs and PMCAs in thirteen different tissues（ skin,brain,gill,liver,muscle,spleen,heart,swim ladder,front intestine, mid-intestine, back intestine, head kidney and hind kidney） by qRT-RCR.Additionally,we studied the changes of the mRNA level in NCXs and PMCAs genes of the gill tissues. Results of blood index showed that both serum osmolality and serum Ca^（2＋）concentrations had a positive correlation with the ambient Ca^（2＋）concentrations. They were significantly different from the control group on day 1 and 4（ P 0. 05）. However,there was not significant difference among Ca^（2＋）treatments on day 7（ P 0. 05）. It indicated that each index changed from high to low level in high calcium treatments,while it changed from low to high in calcium deficiency treatments,and they reached the steady state on day 7. For five blood parameters,the values in treatment groups were higher than that of the control group on day 1 and 4,and they reached the steady state on day 7. Results of tissue distribution showed that NCX2 and NCX3 were highly expressed in the skin,while NCX1,PMCA2 and PMCA4 were highly expressed in the kidney. Results of temporal expression showed that there were distinct differences in NCX1 and PMCA4 with the change of days,while other subunits were not. The NCX1 expression gradually increased in high calcium condition and decreased in low calcium environment wi

关 键 词：花鳗鲡 NCX PMCA 钙稳态 RNA表达量 血液指标 

分 类 号：S917.4[农业科学—水产科学]