Profiling Gene Expression During the Differentiation and Development of the Murine Embryonic Gonad1

Abstract

The application of microarray technology to the study of mammalian organogenesis can provide greater insights into the steps necessary to elicit a functionally competent tissue. To this end, a temporal profile of gene expression was generated with the purpose of identifying changes in gene expression occurring within the developing male and female embryonic gonad. Gonad tissue was collected from mouse embryos at 11.5, 12.5, 14.5, 16.5, and 18.5 days postcoitum (dpc) and relative steady-state levels of mRNA were determined using the Affymetrix MGU74v2 microarray platform. Statistical analysis produced 3693 transcripts exhibiting differential expression during male and/or female gonad development. At 11.5 dpc, the gonad is morphologically indifferent, but at 12.5 dpc, transitions to a male or female phenotype are discernible by the appearance of testicular cords. A number of genes are expressed during this period and many share similar expression profiles in both sexes. As expected, the expression of two well-known sex determination genes, specifically Sry and Sox9, is unique to the testis. Beyond 12.5 dpc, differential gene expression becomes increasingly evident as the male and female tissue morphologically and physiologically diverges. This is evident by two unique waves of transcriptional activity occurring after 14.5 dpc in the male and female. With this study, a large number of transcripts comprising the murine transcriptome can be examined throughout male and female embryonic gonad development and allow for a more complete description of gonad differentiation and development.

Presumptive pre‐sertoli cells express genes involved in cell proliferation and cell signalling during a critical window in early testis differentiation

“…This is not unexpected as pre-Sertoli cells are known to drive the differentiation of the testis immediately after sex determination. Furthermore, these findings confirm observations made by Small et al (2005) who observed a predominance of cell signalling transcripts within the male genital ridge shortly after sex determination. Genes coding for molecules involved in metabolism Genes are classified based on OMIM and SOURCE databases.…”

“…Recently, several large scale transcriptome experiments have been performed to study genital ridge gene expression. Several laboratories have employed subtractive screens or microarray analysis to search for sexually dimorphic gene expression between male and female whole embryonic mouse gonads (Bowles et al, 2000a;Grimmond et al, 2000;Wertz and Herrmann, 2000;Menke and Page, 2002;McClive et al, 2003;Smith et al, 2003;Small et al, 2005). More recently, transgenic mouse models expressing fluorescent proteins driven by various lengths of SRY or Sf1 promoters have been developed and used to compare gene expression of specific cell types within the genital ridge around the time of sex determination and differentiation (Boyer et al, 2004;Nef et al, 2005;Beverdam and Koopman, 2006;Bouma et al, 2006).…”

Screening of genes involved in chromosome segregation during meiosis I: in vitro gene transfer to mouse fetal oocytes

“…[20][21][22][23] Although gene expression profile data for fetal or neonatal ovaries have been provided in mouse and human, and have enabled the analysis of genes involved in the initiation of meiosis, the sex differentiation of the gonad, early meiosis, and ovarian and follicle development, investigations of meiotic prophase I have been lacking. [24][25][26][27][28] In yeast, it is estimated that B150 genes may be meiosis-specific. 29,30 In our current study, we screened 99 genes that are upregulated during prophase in meiosis I.…”

Phylogenetic and expression analysis of the basic helix-loop-helix transcription factor gene family: genomic approach to cellular differentiation

“…High quality mouse RNA samples of at least 5 μg and with a minimum OD260/280 ratio of 1.8 were analyzed as previously described (McLean et al, 2002;Nef et al, 2005;Small et al, 2005). Briefly, RNA was transcribed into cDNA, which was transcribed into biotin labeled RNA.…”

“…Initial analysis of microarray data was performed as previously described (Small et al, 2005). Microarray hybridization data was examined for physical anomalies on the chip and background noise above a value of 3.…”