【摘要】 Nesfatin-1是核连蛋白-2(nucleobindin-2,NUCB2)裂解后产生的一种神经肽。本研究克隆得到斜带石斑鱼两种编码NUCB2(NUCB2a和NUCB2b)基因的c DNA序列(nesfatin-1a和nesfatin-1b),并对它们的表达模式进行了研究。结果表明nesfatin-1a和nesfatin-1b在脑组织中广泛表达。在下丘脑中,两种nesfatin-1在卵巢发育的Ⅰ-Ⅴ时期表达量都比较低,在Ⅵ时期卵巢(退化的卵黄生成阶段的卵母细胞)中的表达量显著升高,揭示nesfatin-1可能参与了卵巢成熟的调控。进一步采用腹腔注射实验研究了两种nesfatin-1对生殖轴的影响。注射nesfatin-1a和nesfatin-1b 15 min和60 min后,下丘脑中促性腺激素释放激素Ⅰ型(Gn RH-Ⅰ)和促性腺激素释放激素Ⅲ型(Gn RH-Ⅲ)的表达量都显著下降;此外,垂体中FSH-β的表达在注射nesfatin-1 60 min后显著下调。这些结果表明nesfatin-1a和nesfatin-1b参与了斜带石斑鱼下丘脑-垂体-性腺轴(HPG)的调控。更多还原

【Abstract】 Nesfatin-1 is the hypothalamus gene encoding nuclear protein-2(nucleobindin-2,NUCB2) produces a neuropeptide after cracking.Studies have shown that nesfatin-1 play an important role in reproductive regulation,in order to study the nesfatin-1 relation with Epinephelus coioides reproductive regulation,in this paper,we have cloned two NUCB2(NUCB2 a and NUCB2b) gene,analyzes NUCB2 coding protein get sequence of amino acids about two nesfatin-1(nesfatin-1a and nesfatin-1b),at the same time,we analysis of the two kinds of nesfatin-1’s signal peptide and mature peptide,found it is consistent with that of vertebrates.Through real-time quantitative PCR technology,we determined the m RNA expression level of Epinephelus coioides nesfatin-1 in different tissues.The expression patterns revealed that,nesfatin-1a and nesfatin-1b widely expressed in the brain tissue,especially nesfatin-1b abundantly expressed in the pituitary,also has the corresponding expression in the gonads.We determined thenesfatin-1 m RNA expression level of hypothalamus during ovary development,the result reveal that,nesfatin-1a and nesfatin-1b in the ovaries Ⅵperiod(degradation of yolk formation stage oocytes) significantly increased,showed that two kinds of nesfatin-1 are likely to participate in the regulation of ovarian mature.The intraperitoneal injection experiment results show that,injecting nesfatin-1a and nesfatin-1b 15 min,both Gn RH1 and Gn RH3 m RNA expression level of the hypothalamus are reduce,but in the pituitary LH-βand FSH-βm RNA expression level has not changed.Injection of nesfatin-1a and nesfatin-1b 60 min,the m RNA expression level of the hypothalamus Gn RH1 and Gn RH3 still suppressed.Injected nesfatin-1a and nesfatin-1b after 60 min,there have no effect change on the expression level of LH-β,but significantly reduced the expression of FSH-β.Through the above study shows nesfatin-1a and nesfatin-1b involved in Epinephelus coioides reproductive regulation.更多还原