Molecular basis of sperm capacitation acrosome reaction and interaction with eggs
A phospholipase C (PLC) which can hydrolyze /sup 14/C-phosphatidylcholine was purified from bull seminal plasma. This PLC has an optimum at pH 7.2 and its PI was about 5.0. The enzyme was inhibited by EDTA, Cd/sup 2 +/, Pb/sup 2 +/, Ni/sup 2 +/, Fe/sup 2 +/, and Zn/sup 2 +/. PLC consists of two subunits one 69,000 and the other 55,000 daltons. The purified PLC was examined for induction of capacitation and acrosome reaction of guinea pig spermatozoa. Sperm were examined for the acrosome reaction 10 min after addition of 3.4 mM Ca/sup 2 +/. Fifty percent of the sperm underwent the acrosome reaction while the control had less than 5% acrosome reacted sperm. The antiserum to the inneracrosomal membrane isolated from sperm was labeled with FITC conjugated goat anti-guinea pig IgG. The conjugated antibody was used to localize sperm antigens. The antigens located on the IAM were only fluoresced when rabbit sperm were treated with methanol and/or MgCl/sub 2/. Therefore anti-IAM antibody did not bind to the sperm plasma membrane. In vivo capacitated rabbit sperm were incubated with anti-IAM antibody (intact IgG and F(ab')/sub 2/ fragments) for 30 min prior to addition of rabbit eggs. After 24 h the eggs were examined for cleavage. The control eggs were fertilized (90%) while the antibody completely inhibited the fertilization of ova in vitro. The eggs incubated with antibody prior to the addition of sperm were still fertilizable. Thus, anti-IAM did not have any noticeable effect on the eggs. It was also shown that antibody inhibited fertilization of zona-free rabbit eggs in vitro as well.
- Research Organization:
- Georgia Univ., Athens (USA)
- OSTI ID:
- 6108594
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
CADMIUM COMPOUNDS
BIOLOGICAL EFFECTS
EDTA
IRON COMPOUNDS
LEAD COMPOUNDS
LIPASE
ENZYME ACTIVITY
PURIFICATION
NICKEL COMPOUNDS
SPERMATOZOA
ANTIGEN-ANTIBODY REACTIONS
ZINC COMPOUNDS
ANTIBODIES
CALCIUM COMPOUNDS
CARBON 14 COMPOUNDS
CATIONS
CATTLE
CHOLINE
EGGS
ENZYME INHIBITORS
GUINEA PIGS
RABBITS
TRACER TECHNIQUES
ALCOHOLS
ALKALINE EARTH METAL COMPOUNDS
AMINES
AMINO ACIDS
AMMONIUM COMPOUNDS
ANIMALS
CARBOXYLESTERASES
CARBOXYLIC ACIDS
CHARGED PARTICLES
CHELATING AGENTS
DOMESTIC ANIMALS
DRUGS
ENZYMES
ESTERASES
GAMETES
GERM CELLS
HYDROLASES
HYDROXY COMPOUNDS
IONS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPOTROPIC FACTORS
MAMMALS
ORGANIC ACIDS
ORGANIC COMPOUNDS
QUATERNARY COMPOUNDS
RODENTS
RUMINANTS
TRANSITION ELEMENT COMPOUNDS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
560301 - Chemicals Metabolism & Toxicology- Cells- (-1987)