上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

Islet1在小鼠睾丸发育过程中的表达研究

陈麒屾，王 路，于 卓，冯立新   

  1. 上海交通大学医学院 医学科学院生殖干细胞研究室, 上海 200025
  • 出版日期:2014-03-28 发布日期:2014-04-02
  • 通讯作者: 冯立新, 电子信箱: fengl3@georgetown.edu; 于 卓, 电子信箱: yuzhuo78@aliyun.com。
  • 作者简介:陈麒屾 （1986—）， 男， 硕士生; 电子信箱: qschen86@gmail.com。
  • 基金资助:

    国家自然科学基金(81000049, 31171409)；国家重大科学研究计划（2012CB966603）

Study on expression of Islet1 in mouse testis development

CHEN Qi-shen, WANG Lu, YU Zhuo, FENG Li-xin   

  1. Laboratory for Germ Cell Research, Institute of Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2014-03-28 Published:2014-04-02
  • Supported by:

    National Natural Science Foundation of China, 81000049, 31171409; Major Scientific Research Program of China, 2012CB966603

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摘要：

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。

关键词: Islet1, 睾丸发育, 生殖细胞, 减数分裂, XY body

Abstract:

Objective To investigate the expression of Islet1 in mouse testis and the process of embryonic organ development in which Islet1 may involved. Methods The RT-PCR was used to detect the time-course expression of Islet1 in mouse testis development and the PCR products were further sequenced and verified. The relative time-course expression quantities and sequences of Islet1 mRNA in mouse testis were measured by the Real-Time PCR. Immunofluorescence staining was then employed to identify Islet1-positive cell types in mouse testis at different time points and subcellular localizations of Islet1-granule were detected by the surface spreading experiment. Results The results of RT-PCR indicated that Islet1 mRNA has transcriptional activation during the entire course of testis development and relative expression quantities of Islet1 mRNA gradually increased with the development of testis tissues in Real-Time PCR results. Immunofluorescence staining showed that the expression of Islet1 was mainly in germ cells. Surface spreading experiments proved that Islet1 localized on the XY body in the nucleus of primary spermatocyte. Conclusion For the first time, we discovered the expression of conservative transcription factor Islet1 in primary spermatocytes of mouse testis and the Islet1 localized on the XY body in the nucleus of primary spermatocyte, which provides new genes for studying the regulation of spermatogonium maintenance and differentiation.

Key words: Islet1, testis development, germ cells, meiosis, XY body