Search

To study the effect of busulfan on the expression and localization of the proliferation and differentiation related proteins PCNA, PLZF, and GATA4 in seminiferous tubules in testis of piglets, in this study, seminiferous tubules of piglets cultured in vitro were selected as the research subjects and divided into control group and experimental group. The experimental group was supplemented with 5 mg/kg of busulfan for 1, 2 and 3 days respectively, and seminiferous tubules were obtained by centrifugation. HE staining was used to detect the morphological changes of chromosomes. Immunohistochemistry, qRT-PCR, and Western blot were used to detect the changes in the expression and localization of PCNA, PLZF, and GATA4. HE staining showed that there were both Sertoli cells and germ cells existing in the seminiferous tubules in all experimental groups. Immunohistochemical studies showed that PCNA protein was localized in the nuclei of germ cells and Sertoli cells in all experimental groups. The staining of PCNA became darker and the expression level increased due to the function of busulfan. In the experimental groups, GATA4 protein was located in the nucleus of Sertoli cell, and there was no significant difference in GATA4 protein staining between the experimental group and the control group. PLZF protein was located in nucleus of germ cells of experimental groups. Compared with the control group, PLZF staining showed no significant difference after 2 days of busulfan treatment, while PLZF staining became darker and expression increased after 1 and 3 days of busulfan treatment. The results of qRT-PCR and Western Blot showed that the mRNA and protein expressions of PCNA and GATA4 in the experimental groups were significantly increased compared with the control group. The expression of PLZF mRNA and protein had no significant differences after 2 days of busulfan treatment. The expression of PLZF mRNA and protein was significantly increased after 1 and 3 days of busulfan treatment. In conclusion, PCNA, PLZF, and GATA4 were expressed in seminiferous tubules of piglets cultured in vitro. The expression levels of PCNA, PLZF, and GATA4 in seminiferous tubules were changed due to the effect of busulfan, suggesting that busulfan damage spermatozoa of porcine testis by affecting the expressions of proteins related to proliferation and differentiation. [ABSTRACT FROM AUTHOR]