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目的探索Islet1在小鼠睾丸中表达情况及其可能参与的生殖细胞发育过程。方法采用RT-PCR技术检测不同发育阶段小鼠睾丸组织中Islet1 mRNA的表达,并对PCR产物进一步测序验证;Real-Time PCR检测不同发育阶段小鼠睾丸中Islet1 mRNA的相对表达量和时序性;免疫荧光染色法进一步检测不同时间点小鼠睾丸中Islet1表达的细胞种类;表面铺展实验检测Islet1在初级精母细胞核中的定位。结果 RT-PCR的结果显示Islet1 mRNA在小鼠睾丸发育整个阶段都有转录活性;Real-Time PCR检测结果发现,随着睾丸组织的发育,Islet1 mRNA的相对表达量逐渐上升;免疫荧光染色结果显示Islet1主要表达在生殖细胞中;表面铺展实验证明了在初级精母细胞的细胞核中,Islet1定位于XY body。结论首次发现胚胎发育中的保守转录因子Islet1在出生后小鼠睾丸的初级精母细胞中存在表达,并在初级精母细胞的细胞核中定位于XY body,这为精原细胞维持和分化的调控研究提供了新的基因。
Objective To explore the expression of Islet1 in mouse testes and its possible involvement in germ cell development. Methods The expression of Islet1 mRNA in testes of different developmental stages was detected by RT-PCR, and the PCR products were further verified by sequencing. The relative expression and the sequence of Islet1 mRNA in testes of different developmental stages were detected by Real-Time PCR. Immunofluorescence staining was used to further detect the cell types of Islet1 expression in mouse testis at different time points. Surface spreading assay was used to detect the localization of Islet1 in primary spermatocyte nuclei. The results of RT-PCR showed that Islet1 mRNA had transcriptional activity throughout the development of mouse testis. Real-Time PCR results showed that the relative expression of Islet1 mRNA increased gradually with the development of testicular tissue. The results of immunofluorescence staining Islet1 is mainly expressed in germ cells; surface spreading experiments demonstrate that Islet1 is localized to the XY body in the nucleus of primary spermatocytes. CONCLUSIONS: It is the first time that the embryonic developmental transcription factor Islet1 is expressed in primary spermatocytes of postnatal mouse testes and localized in the XY body in the nucleus of primary spermatocytes, which regulates the maintenance and differentiation of spermatogonia Research provides new genes.