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Phenotypic analyzes of the epididymis of mice that carry gene mutations that affect the development of the epididymal duct

Grant number: 14/23382-9
Support type:Scholarships abroad - Research
Effective date (Start): March 02, 2015
Effective date (End): September 01, 2015
Field of knowledge:Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
Principal researcher:Raquel Fantin Domeniconi
Grantee:Raquel Fantin Domeniconi
Host: Barry Thomas Hinton
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Research place: University of Virginia (UVa), United States  

Abstract

Spermatozoa acquire ability to recognize or fertilize an egg after passing through the epididymis. Tubular morphogenesis is a fundamental process during the genesis of the epididymis and the mammalian survival depends upon this organ being fully functional. Thus, the elongation and morphogenesis of the Wolffian duct must be highly coordinated with its specialized function of providing an appropriate environment for sperm maturation. Sperm capacitation depends on the modulation of proteins by phosphorylation and dephosphorylation cascades and recently studies have demonstrated that the Src family of kinases plays an important role in the regulation of these events. Despite the normal phosphorylation, sperm from cSrc null mice display a severe reduction in forward motility, and are unable to fertilize in vitro. Preliminary studies conducted by the research group of Dr Hinton suggest that Src plays a role in the development of the epididymis. Therefore, the goal of this project will be to examine the epididymal phenotype of mice that carry a mutation of an inhibitor of SRC, CSK (c-SRC tryrosine kinase). Knockout mice will be generated in which CSK will be removed from the epithelial cells of the initial segment region of the epididymis, during postnatal development. The initial segment has been chosen, because it is critical for male fertility therefore it would be interesting to understand the mechanisms that regulate the development and function of this epididymal region. The analysis of the initial segment phenotype will include: examining the basic histology of the epithelium at the light microscopic level; examining the expression of components of the downstream CSK signaling pathway using Western blot and immunofluorescence for protein levels and RT-PCR and QPCR for mRNA levels and time permits; and performing fertility studies.

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