Genesis Factors Of Chicken Asthenospermia And Association Between Piwill And Chicken Asthenospermia

The continuous declination of fertility in poultry industry caused about10%of chickens been culled. To investigate the factors in the males asthenospermia of cock, an artificial model of chicken asthenospermia was first established, and the semen quality and morphological observation between normal and asthenozoospermia cocks were compared. Afterwards, the expression of Piwill in various kinds of spermatogenic cells were detected by Real-Time quantative PCR. The results were as followed:1. We successfully constructed asthenospermia artificial model in chicken. Compared with the control group, the sperm motility and sperm density of artificial model significantly reduced but the sperm deformity rate increased with increasing treating time. After7d treating with busulfan, seminiferous tubule epithelium began to fall off. Then after treating19d, sperm cell layer and part of the spermatocyte began to fall. However, after25d, there was only spermatogonia layer left. Results of semen smear showed that in manual model, the difference of sperm density and sperm density between control group and treatment group were significantly.2. In self-asthenospermia group, semen volume and sperm density of severe asthenospermia group were significantly lower than that of control group; Sperm abnormality rate of normal group was significantly lower than that of asthenospermia group, while sperm motility was significantly higher than that of asthenospermia group. The difference of sperm density in control group between moderate and severe asthenospermia was significant. 3. The PGCs and SSCs were respectively separated from4.5-day-old and18-day-old chick embryo cells, and their identification results of PAS and SSEA-1staining turned out to be positive. The haploid, diploid and tetraploid cells were obtained from testis cells by flow cytometry sorting. Sperm cells acquired by upstream method. During spermatogenesis, Piwill firstly expressed at lower level in Germ stem cells, and expressed at significantly higher level in spermatogonium cells, lastly had no expression in mature spermatids after the Meiosis. The result of this experiment indicated that the transcripts of Piwill were involved in the regulation of spermatogenesis. The results of asthenozoospermia TaqMan RT-qPCR show that the absolute copy numbers of Piwill in control group was significantly higher than that in naturally asthenozoospermia group and artificial asthenospermia group. The absolute copy numbers of Piwill in treated group decreased with the injection time increasing. The expression levels of Piwill transcript decreased rapidly after7days, and then it reached and maintained at a low expression level at13days.In a word, this experiment had established asthenospermia artificial model in chicken using busulfan.There is a positive correlation between integrit of testicular seminiferous epithelium and semen quality, which is that the more integrity of testicular seminiferous epithelium, the better semen quality. During spermatogenesis, Piwill firstly expressed with lower level in Germ stem cells, and expressed with significantly higher level in the stage of Meiosis, lastly had no expression in mature spermatids after the Meiosis; Piwill played an important role in spermatogenesis, and it had a low mRNA expression in asthenozoospermia cocks, which indicated that Piwill was related to asthenozoospermia.