垂体GnRHR、FHS-β和LH-β基因表达与GnRHR生物信息学研究

Researches of Gene Expression of Pituitary GnRHR,FSH-β and LH-β in Female Rabbits and GnRHR Bioinformatics

为探讨GnRH-A激动剂主动免疫对雌兔垂体GnRHR、FSH-β和LH-βmRNA表达的影响及调节动物生殖功能的分子机理。24只日本大耳白兔分为4组,在EG-I、EG-Ⅱ和EG-Ⅲ兔的颈背侧注射100?g、100?g和50?gGnRH-A抗原,EG-Ⅱ和EG-Ⅲ20d加强注射一次,用荧光定量PCR分析垂体中GnRHR、FSH-β和LH-βmRNA的表达,并测定GnRHR的核苷酸序列。结果表明雌兔垂体中存在GnRHR、FSH-β和LH-β基因。EG-Ⅱ和EG-Ⅲ的GnRHR、FSH-β和LH-βΔΔCt均显著高于EG-Ⅰ(P<0.05)。GnRHR、FSH-β和LH-β的核苷酸序列的同源性分别为98%、100%和94%;GnRHR中α-螺旋占36.63%,理论等电点(pI)5.01,不稳定指数57.08,脂肪指数29.1,疏水性平均值(GRAVY)0.872,表明该蛋白为不稳定的疏水性信号蛋。GnRH-A能明显的影响兔GnRHR、FSH-β和LH-βmRNA在垂体中的表达,而且重复注射会加大这种变化。

The objectives of the study is to explore gene expression of pituitary GnRHR, FSH-15 and LH-β after injecting GnRH -A in female rabbits. Twenty four rabbits（Oryetolagus cunieulus）were divided into EG- Ⅰ , EG - Ⅱ , EG - Ⅲ and control group（each of 6 rabbits）. Rabbits in EG- I , EG- Ⅱ and EG- Ⅲ were subcutaneously injected with 1.0 mL（100 ？ g/mL,100 ？ g/mL and 50 ？ g/mL） GnRH- A antigen respectively. EG- Ⅱ and EG -Ⅲ was re - injected on d 20 at the origin dose. Control group was as blank. The results showed that Gn- RHR, FSH - β and LH - β exist in the pituitary of female rabbits. GnRHR, FSH - β and LH - β AACt in EG - ]I and EG - m were higher than that in EG - I （ P 〈 0.05 ）. The homologies of GnRHR, FSH - 15 and LH - β sequences were respectively 98% , 100% and 94% with those reported in NCBI. In conclusion, GnRH -A at the different dosages may influence obviously the gene expression of GnRHR, FSH - β and LH - β mRNA in rabbit＇s pituitary, re - injection of alarelin had more effects.