The differentiation of mammalian germ cells involves a complex process of morphogenetic changes. To identify the genes which may be involved in thes processes, we have used a strategy based upon the observation that a region of DNA sequence, called the homeobox, is highly conserved in several genes in Drosophila which have been shown to be important in regulating development and differentiation as well as in the genomes of higher organisms. We isolated the mouse homeobox-containing gene Hox-1 4, determined its molecular structure, and examined its tissue, cellular, and temporal specificity of expression at the level of RNA. We will now characterize the Hox-1 4 protein and determine when it is actually translated during spermatogenic differentiation. Antibodies will be generated against the Hox-1 4 protein product. The antibodies will be used to characterize the Hox-1 4 protein and to determine its intracellular localization. We will also examine the possible role of Hox-1 4 in regulating the expression of other homeobox-containing genes expressed in the testis, such as Hox-1 3, Hox-1 5, Hox-2 6 and Hox-5 1. This will initially involve determining the developmental, temporal, and cellular specificity of expression in the testis of these genes and comparing this pattern of expression to that observed in transgenic mice in which Hox-1 4 expression has been altered. Subsequent studies will be directed to determining if the Hox-1 4 protein binds to specific DNA sequences within Hox-1 4 itself and/or to other homeobox-containing genes expressed in the testis. Finally, we will begin to assess the function of Hox-1 4 in the germ line by generating transgenic mouse lines in which we will manipulate the expression of Hox-1 4 by overexpressing Hox-1 4 in germ cells which normally express the gene and by expressing antisense constructs of Hox-1 4 under the direction of spermatogenic-stage specific promoters. These results will provide new insight into the function of homeobox-containing genes during mammalian germ cell development.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD018122-07A1
Application #
3315086
Study Section
Reproductive Biology Study Section (REB)
Project Start
1983-07-01
Project End
1995-02-28
Budget Start
1990-07-01
Budget End
1991-02-28
Support Year
7
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
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Session, D R; Lee, G S; Wolgemuth, D J (2001) Characterization of D1Pas1, a mouse autosomal homologue of the human AZFa region DBY, as a nuclear protein in spermatogenic cells. Fertil Steril 76:804-11
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Pesce, M; Wang, X; Wolgemuth, D J et al. (1998) Differential expression of the Oct-4 transcription factor during mouse germ cell differentiation. Mech Dev 71:89-98
Rhee, K; Brunori, M; Besset, V et al. (1998) Expression and potential role of Fsrg1, a murine bromodomain-containing homologue of the Drosophila gene female sterile homeotic. J Cell Sci 111 ( Pt 23):3541-50
Packer, A I; Crotty, D A; Elwell, V A et al. (1998) Expression of the murine Hoxa4 gene requires both autoregulation and a conserved retinoic acid response element. Development 125:1991-8
Herrada, G; Wolgemuth, D J (1997) The mouse transcription factor Stat4 is expressed in haploid male germ cells and is present in the perinuclear theca of spermatozoa. J Cell Sci 110 ( Pt 14):1543-53
Packer, A I; Elwell, V A; Parnass, J D et al. (1997) N-cadherin protein distribution in normal embryos and in embryos carrying mutations in the homeobox gene Hoxa-4. Int J Dev Biol 41:459-68

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