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Germline Development

Volume 825 of the series Methods in Molecular Biology pp 3-16

Date:

Isolation of Fetal Gonads from Embryos of Timed-Pregnant Mice for Morphological and Molecular Studies

  • Yunmin LiAffiliated withDepartment of Medicine, VA Medical Center, University of California
  • , Teruko TaketoAffiliated withDepartments of Surgery, Biology, and Obstetrics and Gynecology, McGill University, Royal Victoria Hospital
  • , Yun-Fai Chris LauAffiliated withDepartment of Medicine, VA Medical Center, University of California Email author 

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Abstract

Gonadal sex differentiation is an important developmental process, in which a bipotential primordial gonad undergoes two distinct pathways, i.e., testicular and ovarian differentiation, dependent on its genetic sex. Techniques of isolating fetal gonads at various developmental stages are valuable for studies on the molecular events involved in cell-fate determination, sex-specific somatic and germ-cell differentiation and structural organization. Here we describe various procedures for isolation of embryonic gonads at different developmental stages from embryos of timed-pregnant mice. The isolated fetal gonads can be used for a variety of studies, such as organ culture, gene and protein expression. As examples of applications, we describe the immunofluorescence detection of SOX9 expression in gonadal tissue sections and microRNAs profiling/expression in fetal gonads at a critical stage for sex determination.

Key words

Timed-pregnant mice Fetal gonad Mesonephros Testis cords Immunofluorescence microRNAs profiling