| Objective:To explore the variation of c-kit, PI3K(P110α) through studying the expression of the mRNA and protein of c-kit and PI3K(P110α)in bilateral cryptorchidism spermatogonia in different stages of SD rats, and the variation of cell cycle in differentiated type A spermatogonia. Preliminary explore the changes of c-kit gene and downstream signal PI3K (P110α) and cell cycle in SCF/c-kit circuit of differentiated type A spermatogonia by temperature. Methods:The child-bearing age SD rats from the third military medical university were used as the experiment animals, the experiment rats were randomly divided into two groups:the normal control group(without any treatment); the experimental group(the artificial model of bilateral abdominal cryptorchidism). At 1th to 3th weekend after the artificial model of bilateral abdominal cryptorchidism, we observed the expression of c-kit in type A spermatogonia each time point samples with SP immunohistochemistry, The cell cycle of differentiated type A spermatogonia were checked by flow cytometry and the expression of c-kit,PI3K(P110α) mRNA and protein were detected by RT-PCR and Western Blot at the same time. Results:The result of SP immunohistochemistry:The expression of C-kit in the caryon of spermatogonial cells of the normal group, There were plenty of brown particles in the nuclei of spermatogonial cells, and also brown particles could be seen in the nucleus of spermatocytes and acrosome of sperm cells, but staining lighter. The brown particles were not detected in the nuclei of all spermatogenic cells of the negative control. The numbers of spermatogonia were less and less during the experiment time, then finally turned to one single layer. The wall of seminiferous tubule become thin,curved and atrophy, the cavity become larger and interstitial tissue become loose and less. The result of RT-PCR and western blot:l. The expression of c-kit mRNA and protein in differentiated type A spermatogonia from different periods of bilateral abdominal cryptorchidism:c-kit mRNA and protein was highly expressed at 1th weekend and showed decreased tendency as time went on, but higher than the normal group. The c-kit mRNA at 1th weekend,2th weekend,3th weekend was compared respectively with the normal group was significant statistically (P<0.05); the c-kit mRNA at 2th weekend,3th weekend compared was not significant statistically(P>0.05); The c-kit protein at 1th weekend,2th weekend was compared respectively with the normal group was significant statistically (P<0.05). the protein of c-kit at 1th weekend,2th weekend,3th weekend comparison respectively within group showed significant difference (P<0.05). but at 3th weekend with the normal group was not significant statistically(P>0.05).2. The PI3K(P110α)mRNA and protein was highly expressed at 1th weekend and showed decreased tendency as time went on,but higher than normal. The PI3K (P110α)mRNA at 1th weekend,2th weekend,3th weekend was compared respectively with the normal group was significant statistically (P<0.05); The PI3K(P110α) mRNA at 1th weekend 2th weekend,3th weekend comparison respectively within group showed significant difference (P<0.05); The PI3K(P110α) protein at 1th weekend,2th weekend was compared respectively with the normal group was significant statistically (P<0.05),1th weekend,2th weekend,3th weekend was compared respectively within group showed significant difference (P<0.05),3th weekend was compared with the normal group was no significant statistically (P>0.05). The result of cell cycle:Diploid the G0/G1 stage of normal group, the S stage, the G2/M stage and the experimental group was compared respectively was no significant statistically (P>0.05),The percentage of S,G2/M stage were not riseed as time went on. Conclusions:The proliferation signal PI3K(P110α) rised in the proliferating system of SCF/c-kit indicated that the induced proliferation for the rise of temperature may not caused by the SCF/C-KIT pathway.
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