Inhibin a is one of the candidate genes that control the ovulation in poultry. To study the genetic effects of inhibin a on apoptosis and proliferation of goose granulosa cells cultured in vitro, two RNA interference （RNAi） expression vectors, psiRNA-INHal and psiRNA-INHα2, were constructed to knock down inhibin α gene expression. After 48 h of transfection, the efficiency of these two RNAi expression vectors was examined by fluorescence microscopy. Meanwhile, inhibin protein expression levels, apoptosis indexes （AI） and proliferation indexes （PI） of granulosa cells were analyzed by flow cytometry. In addition, the supernatants were collected to assay the concentrations of estrogen （E2） and progesterone （P） by radioimmunoassay. The results showed that the expression level of inhibin a in the RNAi group were decreased 30%--40% than those in the control groups （P 〈0.05） and the apoptosis indexes and proliferation indexes in the RNAi groups were significantly higher than those in the control groups （P 〈0.05）. However, the E2 concentrations in the RNAi groups were lower than those in the control groups （P 〈0.05）. These results indicate that inhibin a has antagonistic effect on granulosa cell apoptosis.
【目的】利用分子标记重构动物之间的亲缘关系。【方法】利用分子标记的同源相同特性,通过对一个随机交配的大群体中任意两个个体在某个标记位点4个等位基因的比较,建立估测群体中任意两个个体之间的亲缘关系的方法(记为Liu模型)。【结果】用蒙特卡罗模拟法(Monte Carlo Simulation)随机模拟4种特殊亲缘关系的群体对Liu模型进行了研究,在4种特定的亲缘关系下,与其它几种模型相比较,Liu模型对个体之间亲缘关系的估测最接近模拟群体的真实值,且估测结果稳定。【结论】建立了估测两个个体之间亲缘关系的Liu模型,通过比较分析证明了Liu模型是目前估测效果最好的模型。
Inhibin was purified from swine ovarian fluid by deposition and column deposition.The purified inhibin was diluted with ureasodiumacetate,and then was mixed with Freund adjuvant with same volume.Each of the twenty goats was given subcutaneous injections of the purified inhibin 2 mg at neck.The goats were given boosterinjections of the inhibin 1 mg that was mixed with ureasodiumacetate and Freund incomplete adjuvant after 30 and 50 days.Five goats in control group were given admixture of urea and sodium acetate and adjuvant at the same time.The goats were injected with prostaglandin F2a 005 mg at 21 d and 31 d after the last immunization.The blood samples were collected at 30 d,51 d and 60 d after the first immunization and before immunization.The plasma FSH concentrates was assayed with RIA and the inhibin antibody titer was assayed with agar plate diffusion method.The plasma antibody titer was 1∶16256 at 60 d after first immunization.The plasma FSH concentrates of the immunized goats was higher than those of the control group(202 vs.12 mIU/ml,PZ<005).The number of follicle and the number of corpora lutea of the immunized group were as twice as those of the control(1931 vs.1223 and 225 vs.110,respectively).It indicated that inhibin purified from swine ovarian fluid could induce goat produce high ovulation rate and it can improve reproductive traits of goat.
