The differentiation of specific cell types in mammals is mediated by differential gene expression. To understand the regulation of the development and differentiation of a particular cell type, it is important to understand what differential gene expression occurs in that cell type and how this gene expression is regulated. For the last 12 years we have been studying one gene (phosphoglycerate kinase-2 or Pgk-2) that is specifically expressed in spermatogenic cells in mammals. In that time, we have learned a great deal about the expression, regulation, structure, function and evolution of this gene. In this application, we propose additional experiments to help us further understand the how the spermatogenesis-specific expression of the Pgk-2 is regulated at the level of transcription. We believe that by learning more about how the expression of this single gene is regulated in these cells, we will gain insight into how many of the genes that are differentially expressed in spermatogenic cells might be regulated. In this way, we hope to contribute to a better understanding of how the process of spermatogenesis is, itself, regulated. Our emphasis in this application is to study the developmental order of molecular events that predispose the initiation of transcription of the Pgk-2 gene in spermatogenic cells, and also to follow the """"""""reprogramming"""""""" of these events following cessation of Pgk-2 transcription.
The specific aims of this application include an investigation of the occurrence of protein-DNA interactions in the promoter region of the Pgk-2 gene in spermatogenic cells in vivo, a molecular characterization of testis-specific nuclear transcription factors that are critically involved in the tissue-specific regulation of transcription of the Pgk-2 gene, and a developmental study of changes in chromatin structure, association of the Pgk-2 gene with the nuclear matrix, and timing of its replication that predispose and follow active transcription of this gene. We believe that by learning more about the order of the various molecular events that regulate Pgk-2 transcription, we will gain a much better understanding of die overall mechanism of gene regulation in spermatogenic cells. This could provide valuable insight into avenues of treatment for defective gene expression during spermatogenesis, and/or other genetic manipulations involving these cells.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD023126-09
Application #
2403158
Study Section
Reproductive Biology Study Section (REB)
Project Start
1987-04-01
Project End
1998-11-30
Budget Start
1997-12-01
Budget End
1998-11-30
Support Year
9
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Southwest Foundation for Biomedical Research
Department
Type
DUNS #
City
San Antonio
State
TX
Country
United States
Zip Code
78245
Wang, P Jeremy; Page, David C; McCarrey, John R (2005) Differential expression of sex-linked and autosomal germ-cell-specific genes during spermatogenesis in the mouse. Hum Mol Genet 14:2911-8
Wykes, Susan M; Krawetz, Stephen A (2003) Separation of spermatogenic cells from adult transgenic mouse testes using unit-gravity sedimentation. Mol Biotechnol 25:131-8
Wykes, Susan M; Krawetz, Stephen A (2003) Conservation of the PRM1 --> PRM2 --> TNP2 domain. DNA Seq 14:359-67
Wykes, Susan M; Krawetz, Stephen A (2003) The structural organization of sperm chromatin. J Biol Chem 278:29471-7
McCarrey, John R; Watson, Cathy; Atencio, Julia et al. (2002) X-chromosome inactivation during spermatogenesis is regulated by an Xist/Tsix-independent mechanism in the mouse. Genesis 34:257-66
Klaus, A V; McCarrey, J R; Farkas, A et al. (2001) Changes in DNA loop domain structure during spermatogenesis and embryogenesis in the Syrian golden hamster. Biol Reprod 64:1297-306
Kramer, J A; McCarrey, J R; Djakiew, D et al. (2000) Human spermatogenesis as a model to examine gene potentiation. Mol Reprod Dev 56:254-8
Davis, T L; Yang, G J; McCarrey, J R et al. (2000) The H19 methylation imprint is erased and re-established differentially on the parental alleles during male germ cell development. Hum Mol Genet 9:2885-94
Krawetz, S A; Kramer, J A; McCarrey, J R (1999) Reprogramming the male gamete genome: a window to successful gene therapy. Gene 234:1-9
McCarrey, J R; O'Brien, D A; Skinner, M K (1999) Construction and preliminary characterization of a series of mouse and rat testis cDNA libraries. J Androl 20:635-9

Showing the most recent 10 out of 25 publications