Methylation of the Sox9 and Oct4 promoters and its correlation with gene expression during testicular development in the laboratory mouse.

Mamta Pamnani,Puja Sinha,Seema Nara,Alka Singh,Manisha Sachan

doi:10.1590/1678-4685-gmb-2015-0172

Abstract

Sox9 and Oct4 are two important regulatory factors involved in mammalian development. Sox9, a member of the group E Sox transcription factor family, has a crucial role in the development of the genitourinary system, while Oct4, commonly known as octamer binding transcription factor 4, belongs to class V of the transcription family. The expression of these two proteins exhibits a dynamic pattern with regard to their expression sites and levels. The aim of this study was to investigate the role of de novo methylation in the regulation of the tissue- and site-specific expression of these proteins. The dynamics of the de novo methylation of 15 CpGs and six CpGs in Sox9 and Oct4 respectively, was studied with sodium bisulfite genomic DNA sequencing in mouse testis at different developmental stages. Consistent methylation of three CpGs was observed in adult ovary in which the expression of Sox9 was feeble, while the level of methylation in somatic tissue was greater in Oct4 compared to germinal tissue. The promoter-chromatin status of Sox9 was also studied with a chromatin immune-precipitation assay.

Highlights

Sox9 and Oct4 are two developmentally important genes that show differential spatial and temporal expression
Consistent methylation of three CpGs was observed in adult ovary in which the expression of Sox9 was feeble, while the level of methylation in somatic tissue was greater in Oct4 compared to germinal tissue
Since methylation can affect gene expression in various tissues (Sachan et al, 2006; Pamnani et al, 2014), in this study, we investigated the status of de novo methylation and expression pattern in the promoter region of mouse Sox9 and Oct4 gene during development

Summary

Introduction

Sox and Oct are two developmentally important genes that show differential spatial and temporal expression. SRY (sex determining region Y) box 9 (Sox9) , located on chromosome 11 possesses a Sry-box which is the first gene to be expressed shortly after Sry gene expression (Chaboissier et al, 2004; Koopman 2005; Vidal et al, 2001). Sox expression was identified in the genital ridges of male and female sex gonads in 11.5 days post-coitum (dpc) embryos (Chaboissier et al, 2004; Sekido et al, 2004). After day 12.5, expression of Sox is totally restricted to Sertoli cells and maintained until the adult stage (Kobayashi et al, 2005; Koopman, 2005). RT-PCR and immunoblot identified Sox expression in mature adult testis but not in adult ovary (Kent et al, 1996).

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