The long-term ojective of these studies is to understand how the synthesis of specific temporally expressed proteins is regulated in the mammalian testis. Emphasis will be placed upon establishing the function of a unique testicular a-tubulin and defining the cis-acting regulatory sequences in the genomic DNA of this tubulin and several other haploid gene products including the two protamines. Additional studies will explain how coordinate expression of genes in pre-meiotic, meiotic and post-meiotic testicular cells is controlled. Specific projects to be undertaken include: 1) determining the localization in testicular cells by immunofluorescence of the unique post-meiotically synthesized mouse a-tubulin. 2) determining whether a common DNA enhancer sequence regulates the synthesis of the protamine, a-tubulin, and other coordinately expressed haploid genes. 3) identifying specific cDNA and genomic clones for pre-meiotic and meiotic testicular cell-types to allow a direct comparison of genomic regulatory signals to be made between different classes of temporally regulated genes. 4) establishing the relationship between DNA methylation and the gene transcription and nuclear geome condensation with our growing collection of cell-type specific probes. These studies on the molecular basis for gene regulation in the testis will enhance our knowledge of basic control mechanisms during spermatagenesis, provide necessary probes to understand molecular causes of infertility, and provide superior approaches to fertility control.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM029224-11
Application #
3276768
Study Section
Reproductive Biology Study Section (REB)
Project Start
1980-09-01
Project End
1991-08-31
Budget Start
1990-09-01
Budget End
1991-08-31
Support Year
11
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Tufts University
Department
Type
Schools of Arts and Sciences
DUNS #
073134835
City
Medford
State
MA
Country
United States
Zip Code
02155
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Hermo, L; Oko, R; Hecht, N B (1991) Differential post-translational modifications of microtubules in cells of the seminiferous epithelium of the rat: a light and electron microscope immunocytochemical study. Anat Rec 229:31-50
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Oko, R; Hermo, L; Hecht, N B (1991) Distribution of actin isoforms within cells of the seminiferous epithelium of the rat testis: evidence for a muscle form of actin in spermatids. Anat Rec 231:63-81
Johnson, P A; Bunick, D; Hecht, N B (1991) Protein binding regions in the mouse and rat protamine-2 genes. Biol Reprod 44:127-34
Kwon, Y K; Hecht, N B (1991) Cytoplasmic protein binding to highly conserved sequences in the 3' untranslated region of mouse protamine 2 mRNA, a translationally regulated transcript of male germ cells. Proc Natl Acad Sci U S A 88:3584-8
Alcivar, A A; Trasler, J M; Hake, L E et al. (1991) DNA methylation and expression of the genes coding for lactate dehydrogenases A and C during rodent spermatogenesis. Biol Reprod 44:527-35
Morales, C R; Kwon, Y K; Hecht, N B (1991) Cytoplasmic localization during storage and translation of the mRNAs of transition protein 1 and protamine 1, two translationally regulated transcripts of the mammalian testis. J Cell Sci 100 ( Pt 1):119-31
Bunick, D; Johnson, P A; Johnson, T R et al. (1990) Transcription of the testis-specific mouse protamine 2 gene in a homologous in vitro transcription system. Proc Natl Acad Sci U S A 87:891-5

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