Abstract
Spata34 is a testis-specific-expressed gene which exerts diverse functions in testis development. This study intends to examine the expression profiles of Spata34 in postnatal rat testis, and explore its potential roles in cell proliferation in vitro. We found that the mRNA and protein expression levels of Spata34 were developmentally upregulated in rat testes during the early 1–7 postnatal weeks using real-time polymerase chain reaction and western blotting. Immunohistochemical results indicated that Spata34 protein was mainly detected in the nuclear and cytoplasm of spermatocytes and round spermatids. The possible function of Spata34 in cellular proliferation was analyzed using cell counting kit, colony formation and flow cytometry assays. Our results showed that overexpression of Spata34 in multipotent adult germline stem cell lines (maGSC129SV) cells markedly facilitated cell proliferation with a large increase in cell numbers in S phase of cell cycle. While knockdown of Spata34 expression by specific siRNA suppressed the growth of maGSC129SV cells and triggered cell-cycle arrest at G1/S phase transition, which was related to the elevation of p21 and p27 and decrease of Cyclin D1 and Cyclin D-dependent kinase 4. Altogether, our results indicated that the Spata34 gene evokes unique expression patterns during postnatal development of the rat testis, and for the first time, unravels the function of Spata34 on regulating cell-cycle progress through p21 and p27 pathway.
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Acknowledgements
This study was supported by the National Natural Science Foundation of China (Nos. 31571277, 30971570). The authors would like to thank Dr Zhang QianJun from Central South University (China) for kindly providing the GC-1 cell lines and maGSC129SV cell lines used in the experiment.
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Conceived and designed the experiments: DN and ZL; performed the experiments: HL and YL; contributed reagents/materials/analysis tools: DN, HL, YL and ZL; wrote the paper: DN; completed calculations and participated in the interpretation of the results: HL and ZL.
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All animal handling complied with the requirements of the directory of the Ethical Treatment of Experimental Animals of China.
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Nie, D., Li, H., Liu, Y. et al. Role of Spata34 in cell proliferation and its expression pattern in postnatal development of rat testis. Mol Biol Rep 45, 2697–2705 (2018). https://doi.org/10.1007/s11033-018-4439-6
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DOI: https://doi.org/10.1007/s11033-018-4439-6