New Candidate Genes Identified for Controlling Mouse Gonadal Sex Determination and the Early Stages of Granulosa and Sertoli Cell Differentiation1

Abstract

Mammalian gonadal sex-determining (GSD) genes are expressed in a unique population of somatic cells that differentiate into granulosa cells in XX gonads or Sertoli cells in XY gonads. The ability to efficiently isolate these somatic support cells (SSCs) during the earliest stages of gonad development would facilitate identifying 1) new candidate GSD genes that may be involved in cases of unexplained abnormal gonad development and 2) genes involved in the earliest stages of granulosa and Sertoli cell differentiation. We report the development of a unique mouse carrying two transgenes that allow XX and XY mice to be distinguished as early as Embryonic Day 11.5 (E11.5) and allow SSCs to be isolated from undifferentiated (E11.5) and early differentiated (E12.5) fetal gonads. The Mouse Genome 430v2.0 GeneChip (Affymetrix) was used to identify transcripts exhibiting a sexual dimorphic expression pattern in XX and XY isolated SSCs. The analysis revealed previously unidentified sexually dimorphic transcripts, including low-level expressed genes such as Sry, a gene not identified in other microarray studies. Multigene real-time PCR analysis of 57 genes verified that 53 were expressed in fetal gonads in a sexually dimorphic pattern, and whole-mount in situ hybridization analysis verified 4930563E18Rik, Pld1, and Sprr2d are expressed in XX gonads, and Fbln2, Ppargc1a, and Scrn1 are expressed in XY gonads. Taken together, the data provide a comprehensive resource for the spatial-temporal expression pattern of genes that are part of the genetic network underlying the early stages of mammalian fetal gonadal development, including the development of granulosa and Sertoli cells.

Uncovering Gene Regulatory Networks During Mouse Fetal Germ Cell Development

“…In this regard, the outcome of our experiment is very similar to that of previous screens dedicated to gene expression in the somatic compartment of developing gonads [24][25][26]. Perhaps more importantly, this dataset could shed light on the influence of germ cells over somatic development.…”

“…A number of gene expression screens, notably several microarray experiments, have been conducted with fetal gonads [23]. Despite the large amounts of data generated, 1 most of those studies were focused on expression programs underlying sex determination in somatic cells [24][25][26] or were based on whole gonads, in which it is not possible to distinguish between somatic and germ cell gene expression [27,28]. Two studies used isolated mouse germ cells to compare gene expression in pre-versus postmigratory PGCs [28] or embryo-derived stem cells versus PGCs [29].…”

Heterogeneity in sexual bipotentiality and plasticity of granulosa cells in developing mouse ovaries

“…1I). Moreover, some of these cells appeared to be positive for SPRR2d, another early pre-granulosa cell marker (Lee et al, 2009;Bouma et al, 2010) (supplementary material Fig. S2B).…”

Purification and Transcriptomic Analysis of Mouse Fetal Leydig Cells Reveals Candidate Genes for Specification of Gonadal Steroidogenic Cells1

“…Previous transcriptomic studies aimed at identifying genes important for development of the fetal gonads in mice, or for establishing the molecular signatures of the component cell lineages, have been performed using microarrays [12][13][14][15][16]. While this method reveals the expression dynamics of thousands of genes simultaneously, it is limited by the incomplete representation of genes on the array and also by the relatively low sensitivity and dynamic range offered [17].…”