齐口裂腹鱼和鲤鱼H-FABP基因的克隆及其表达谱

心型脂肪酸结合蛋白(heart fatty acid binding protein,H-FABP)的水平与影响肉质性状的肌内脂肪含量有关,鱼类H-FABP的表达水平对其肌内脂肪含量是否相关仍未见报道.本研究获得齐口裂腹鱼和鲤鱼心脏型脂肪酸结合蛋白基因序列,利用半定量RT-PCR分析其表达特性并测定肌内脂肪含量,比较H-FABP基因在不同生活环境的2种鲤科鱼肌内脂肪沉积中的作用.结果显示,齐口裂腹鱼和鲤鱼H-FABP基因的ORF为402 bp,编码133个氨基酸,它们的氨基酸序列相同,与人、猪、小鼠、斑马鱼、大西洋鲑、虹鳟等的同源性为71.3%～90%;H-FABP基因在2种鲤科鱼的心、肌肉、脂肪、肝、脑、脾、肾和鳃等组织中均有表达,肝中的表达量显著高于其它组织(P<0.05),H-FABP基因的肌肉表达谱在齐口裂腹鱼和鲤鱼中存在明显差异:齐口裂腹鱼中的表达随生长发育呈上升趋势,在大体重鱼(500 g)中的表达显著高于小体重鱼(P<0.05),其表达与肌内脂肪含量呈显著正相关(R=0.370,P<0.05);H-FABP基因在鲤鱼生长发育中呈下降趋势,而小体重鱼(50～60 g)中的表达显著高于其它大体重鱼(P<0.05),其表达与肌内脂肪含量呈显著负相关(R=-7.083,P<0.01).据此推测,齐口裂腹鱼和鲤鱼肌肉组织H-FABP基因表达与肌内脂肪关联性的差异可能与2种鱼的生活环境不同有关.

Study on HSP70 Gene Expression in Different Tissue of Cyprinus carpio

[ Objective] The aim of this study was to investigate whether HSP70 can be used as a stress monitoring indicator in Cypnnus carpio breeding. [Method] Based on HSP70 sequence of Cyprinus carpio （AY120894）, one pair of primers was designed and synthesized, while the total RNA of liver tissues in Cyprinus carpio was extracted. Some cDNA fragments of Cyprinus carpio HSP70 were cloned by RT-PCR, and its differential expression in various tissues such as heart, intestine, mucus, gonad, swim bladder, gill and fin in Cyprinus carpio was also studied. [Result] The cDNA sequence of 480 bp was obtained from Cypdnus carpio HSPTO gene by RT-PCR amplification. Homology comparison between the deduced amino acid sequence after sequencing and that of other types of fish showed that the homology among Cyprinus carpio, Danio rerio, Ohcorhynehus mylciss, Paralichthys olivaceus, Xiphophoorus maculates and Carassius auratus was 96%, 98%, 98%, 96%, 98% and 96% respectively. The expression of HSP70 was detected in eight tissues of Cypnnus carpio. The expression was the highest in heart, followed by swim bladder and fin, but there was no significant difference between them （ P 〉 0.05 ）. There was no significant difference among the ex- pression in three tissues of intestine, mucus and fat （ P〉0.05）, but their expression was significantly higher than those in gonad and gill （ P〈 0.05）. [ Conclusion] HSPTO gene expression is a suitable criterion for monitoring the stress degree, stress capacity and healthy conditions in Cyprinus carpio breeding.

添加抗氧化物维生素E对人类胚胎体外发育的影响

目的:探讨抗氧化物维生素E添加与人类胚胎体外发育潜力之间的关系。方法:选择2009年10月至2010年5月在我院进行常规体外受精-胚胎移植和卵胞浆内单精子注射的不孕患者106例,116个周期,共432个正常受精2个原核(2PN)来源的废弃胚胎,随机分为对照组和维生素E组,在维生素E组受精后第3天(D3)废弃胚胎的培养液中添加100μmol维生素E,对照组不添加,观察两组囊胚的形成情况,并通过荧光染料Hoechst33342对胚泡进行染色,检测囊胚的总细胞数。结果:维生素E组的早期囊胚、扩张囊胚、孵化囊胚发育率均高于对照组,两组比较差异有统计学意义(P<0.05);维生素E组孵化囊胚细胞总数(80.42±0.60)%高于对照组(76.11±0.82)%,差异有统计学意义(t=-7.365,P<0.05)。结论:添加维生素E能提高人类胚胎的体外发育率和胚胎质量。