The Observation Of The Level Of Cytokines And The Histological Change Of The Hypothalamus-pituitary-testicle Axle Of Attractin Gene Knockout Male Mice

Experiment I The change of the cytokines level of male Attractin gene knockout miceObjective To examine the influence to the Cytokines level of Attractin gene knockout male mice.Methods Male mature Attractin gene knockout homozygote mice and wild type mice(C3H/hej)were divided into 2 groups: the Atrn knockout group and the control group. Each group included 12 mice. Blood samples were collected from angular vein of mice. The level of IL-2,IL-6 and TNF-αwas determined with ELISA. The results would be compared between groups.Results For the Atrn knockout group, the level of IL-2 was lower than that of the control group (P<0.05), while the level of IL-6 and TNF-αwas higher (P<0.05).Conclusion Knocking out Atrn gene not only can down-regulate the level of IL-2 of male mice, but also can up-regulate the level of IL-6 and TNF-αof male mice. The mechanism of this phenomenon needs further study. ExperimentⅡThe histological change of the hypothalamus-pituitary-testicle axle of Attractin gene or Mahoganoid (Md) gene knockout male miceObjective To examine the influence to the histological HE slices of the hypothalamus-pituitary-testicle axle of Attractin gene or Mahoganoid (Md) gene knockout male mice.Methods Male mature Attractin gene knockout homozygote mice, Male mature Md gene gene knockout homozygote mice and Male mature wild type mice(C3H/hej)were divided into 3 groups: the Atrn knockout group, the Md knockout group and the control group. Each group included 6 mice. The hypothalamus, the pituitary gland, the testicle and the epididymis of each group mice were taken out, fixed, embeded with wax and maken into HE slices.Results 1. Obvious spongy degeneration was observed in the hypothalamus and the pituitary gland of the male mature Atrn gene gene knockout homozygote mice or male mature Md gene gene knockout homozygote mice. 2. There was no obvious change in the HE slices of the testicle and the epididymis.Conclusion Obvious spongy degeneration was observed in the hypothalamus and the pituitary gland of the male mature Atrn gene or Md gene knockout homozygote mice. The HE slices of the testicle and the epididymis had no visible change.ExperimentⅢThe observation of the seminal fluid parameter of Attractin (Atrn) gene knockout male mice Objective To examine the influence to the seminal fluid parameter of Attractin (Atrn) gene knockout male mice.Methods Male mature Atrn gene knockout homozygote mice and wild type mice(C3H/hej)were divided into 2 groups: the Atrn knockout group and the control group. Each group included 6 mice. After cervical vertebra dislocation execution, the bilateral testicle, epididymis and seminal vesicle were taken out immediately, separated from the fat and the fascia, attractted dryly by the filter paper. One side cauda epididymidis and the partial deferent duct were put into the 0.3mlF-10 nutrient fluid which was warmed in advance. Then these tissues were cut into fragments. Spermato-suspension was taken as more as possible after fostering 5~10minutes. The tissues fragments were abandoned. The volume was recorded. The seminal fluid parameter was examined, and the obtained data was eacuted a statistics analysis.Results The sperm speed of the Atrn knockout group was lower than that of the control group (P<0.05). Compared with the control group, there was no obvious difference in the sperm density, sperm motility rate, curling tail rate of spermatozoon in the osmotic pressure swell experiment, sperm activity ratio, organ coefficient of testicle, epididymis and seminal vesicle of the Atrn knockout group.Conclusion Knocking out Atrn gene may play an important role in the process of spermatogenisis but its specific mechanism remains to be clarified.