The formation of flagella and the synthesis of proacrosin will be investigated in primary cultures of mouse round spermatids. Initial experiments are designed to determine whether spermatids that do not make flagella in culture lack the centrioles necessary for axonemal nucleation. The requirements of protein, RNA, and tubulin biosynthesis for flagella formation will also be examined. The length of time for cell culture will be increased to ascertain the extent of tail maturation that can be obtained in vitro. Antibodies to the major fibrous sheath protein will be used to determine whether this component appears in the newly-formed flagella. The synthesis of proacrosin will also be examined to determine whether proacrosin contains N-linked oligosaccharides and whether these carbohydrate side chains are processed to the """"""""complex"""""""" type. The carbohydrate side chains will be analyzed for mannose-6-phosphate groups. If these residues are found, it suggests that proacrosin is directed to the acrosome by the same phosphomannosyl receptor system that transports lysosomal hydrolases to the lysosome. The experiments of this proposal will provide the bases for long-term biochemical studies of mammalian spermiogenesis.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Unknown (R23)
Project #
5R23HD020736-02
Application #
3448265
Study Section
Reproductive Biology Study Section (REB)
Project Start
1984-12-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Carrera, A; Gerton, G L; Moss, S B (1994) The major fibrous sheath polypeptide of mouse sperm: structural and functional similarities to the A-kinase anchoring proteins. Dev Biol 165:272-84
Anakwe, O O; Gerton, G L (1990) Acrosome biogenesis begins during meiosis: evidence from the synthesis and distribution of an acrosomal glycoprotein, acrogranin, during guinea pig spermatogenesis. Biol Reprod 42:317-28
Joshi, M S; Anakwe, O O; Gerton, G L (1990) Preparation and short-term culture of enriched populations of guinea pig spermatocytes and spermatids. J Androl 11:120-30
Kopf, G S (1990) Zona pellucida-mediated signal transduction in mammalian spermatozoa. J Reprod Fertil Suppl 42:33-49
Kopf, G S (1989) Mechanisms of signal transduction in mouse spermatozoa. Ann N Y Acad Sci 564:289-302
O'Brien, D A; Gerton, G L; Eddy, E M (1988) Acrosomal constituents identified with a monoclonal antibody are modified during late spermiogenesis in the mouse. Biol Reprod 38:955-67
Arboleda, C E; Gerton, G L (1988) Proacrosin/acrosin during guinea pig spermatogenesis. Dev Biol 125:217-25
Gerton, G L; O'Brien, D A; Eddy, E M (1988) Antigens recognized by monoclonal antibody to mouse acrosomal components differ in guinea pig spermatogenic cells and sperm. Biol Reprod 39:431-41
Adekunle, A O; Arboleda, C E; Zervos, P H et al. (1987) Purification and initial characterization of guinea pig testicular acrosin. Biol Reprod 37:201-10
Arboleda, C E; Gerton, G L (1987) Studies of three major proteases associated with guinea pig sperm acrosomes. J Exp Zool 244:277-87

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