Abstract
REGULATION OF NT3 IN THE EMBRYONIC GONAD AT THE TIME OF TESTIS DIFFERENTIATION AND IDENTIFICATION OF NEW CANDIDATE GENES
Biology of reproduction, Vol.77(Suppl_1), pp.133-133
07/01/2007
Abstract
Nt3 plays a significant role in male gonadal development. It is required for mesonephric cell migration into the testis for cord formation. The expression profiles for Nt3 are consistent with those expected for a gene directly regulated by SRY, therefore it is being studied as a candidate. Nt3 is highly conserved between species and potential SRY binding HMG box elements were identified in the Nt3 promoter. Promoter constructs with luciferase reporters were made for mouse and rat Nt3 promoters. Effects of mutations to the putative SRY binding elements was monitored in transfection assays. Primary Sertoli cell cultures from postnatal day 20 rats were used for transfection. Primary embryonic day 13 testis cells were also cultured and used in transfection assays. The mutated sites in the mouse and rat Nt3 promoters were found to be important for full promoter activity in these cells, and mutated promoters had reduced reporter activity. SOX9 is also important for sex determination and like SRY, contains an HMG domain that is able to bind DNA at highly similar binding elements. Therefore the potential role of both SRY and SOX9 in regulation of the Nt3 promoter were investigated. The potential for SRY and SOX9 to exert an effect on the Nt3 promoter was examined in co-transfection assays using Sry and Sox9 expression vectors. Initial data shows that SRY can exert an effect in transfection assays, however we are in the process of determining if SOX9 has the same or a greater effect on the Nt3 promoter. RNAi is being used to determine if there is a morphological effect or change in Nt3 expression when Sry or Sox9 are silenced at the time of initiation of gonadal differentiation. Other candidate genes for involvement in sex determination and regulation downstream of Sry were identified in a microarray analysis of embryonic gonads in the rat. A functional role for select testis development candidates is now being investigated. These studies have helped clarify the regulation of Nt3 in the gonad during sex determination and identified new candidates whose function and regulation will be investigated in a manor similar to studies of Nt3.
Metrics
1 Record Views
Details
- Title
- REGULATION OF NT3 IN THE EMBRYONIC GONAD AT THE TIME OF TESTIS DIFFERENTIATION AND IDENTIFICATION OF NEW CANDIDATE GENES
- Creators
- Tracy Clement - Washington State UniversityMatthew Anway - Washington State UniversityMichael Skinner - Washington State University
- Publication Details
- Biology of reproduction, Vol.77(Suppl_1), pp.133-133
- Academic Unit
- Biological Sciences, School of
- Identifiers
- 99901080302401842
- Language
- English
- Resource Type
- Abstract