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Creatine kinase isoenzymes in spermatozoa
Fertilization ( 4 5 3 - 4 6 0 )
454
453
CREATINE KINASE ISOENZYMES IN SPERMATOZOA H . M . E p p e n b e r g e r , H.Moser, B . Z u r b r i g g e n , T.Wallimann. Dept. of Cell Biology, E.T.H., CH-8093 Zurich, Switzerland. Two isoforms of creatine Kinase, the brain type BB-CK and the mitochondrialbound MiMi-CK, as well as adenylate kinase were identified in washed spermatozoa from chicken and man by cellulose polyacetate electrephoresis and immuneblots. BB-CK was localized by indirect immunofluorescence staining within the sperm tail but not in the head portion. MiMi-CK is confined to the midpieee region rich in mitoehondria. In contrast to chicken, seminal plasma from man was also found to contain considerable amounts of BB-CK. Total oreatine content of spermatozoa [8-15 mM] and seminal plasma [3.8 + 0.4 mM) as well as preliminary experiments with metabolic bloKKers indicate a dependence of sperm m o t ~ lity on CK and phosphoryl-ereatine [CPJ. The presence of two CK isoforms located in different "compartments" of spermatozoa suggests a CP-shuttle in sperms similar to that described for crossstriated muscle.
PURIFICATION AND IMMUNOCYTOCHEMICAL LOCALIZATION OF THE VlTELLINE COAT-LYSIN FROM THE ACROSOMAL VESICLES OF ABALONE SPERM. K.Haino-Fukushima and N.Usui Department of Biology,Tokyo Metropolitan University, Setagaya, Tokyo 158, Japan. Spermatozoa of the Japanease abalone Haliotis discus were obtained by exposing the animals to UV-irradiated seawater and induced to undergo the acrosome reaction in high-calcium seawater. The soluble components released from the aerosomes showed a potent lyric activity on the vitelline eoat(VC). From the components, the VC lysin was purified by salting-in, preparative polyacrylamide gel electrophoresis(PAGE) and high performance liquid chromatography. Its molecular weight was estimated to be 15,500 by SDSPAGE and its isoeleetrie point was pH 9.6, suggesting that the VC lysin existed as a basic monomerie molecule. In addition, against the purified VC lysin, antibody was raised in a rabbit. Using it, the antigenic sites were labeled on the thin sections of the acrosomal vesicles, by applying the protein A-gold technique. The anti-15.5K lysin antibody was well localized at the posterior half of the acrosomal vesicle.
183S
454
453
CREATINE KINASE ISOENZYMES IN SPERMATOZOA H . M . E p p e n b e r g e r , H.Moser, B . Z u r b r i g g e n , T.Wallimann. Dept. of Cell Biology, E.T.H., CH-8093 Zurich, Switzerland. Two isoforms of creatine Kinase, the brain type BB-CK and the mitochondrialbound MiMi-CK, as well as adenylate kinase were identified in washed spermatozoa from chicken and man by cellulose polyacetate electrephoresis and immuneblots. BB-CK was localized by indirect immunofluorescence staining within the sperm tail but not in the head portion. MiMi-CK is confined to the midpieee region rich in mitoehondria. In contrast to chicken, seminal plasma from man was also found to contain considerable amounts of BB-CK. Total oreatine content of spermatozoa [8-15 mM] and seminal plasma [3.8 + 0.4 mM) as well as preliminary experiments with metabolic bloKKers indicate a dependence of sperm m o t ~ lity on CK and phosphoryl-ereatine [CPJ. The presence of two CK isoforms located in different "compartments" of spermatozoa suggests a CP-shuttle in sperms similar to that described for crossstriated muscle.
PURIFICATION AND IMMUNOCYTOCHEMICAL LOCALIZATION OF THE VlTELLINE COAT-LYSIN FROM THE ACROSOMAL VESICLES OF ABALONE SPERM. K.Haino-Fukushima and N.Usui Department of Biology,Tokyo Metropolitan University, Setagaya, Tokyo 158, Japan. Spermatozoa of the Japanease abalone Haliotis discus were obtained by exposing the animals to UV-irradiated seawater and induced to undergo the acrosome reaction in high-calcium seawater. The soluble components released from the aerosomes showed a potent lyric activity on the vitelline eoat(VC). From the components, the VC lysin was purified by salting-in, preparative polyacrylamide gel electrophoresis(PAGE) and high performance liquid chromatography. Its molecular weight was estimated to be 15,500 by SDSPAGE and its isoeleetrie point was pH 9.6, suggesting that the VC lysin existed as a basic monomerie molecule. In addition, against the purified VC lysin, antibody was raised in a rabbit. Using it, the antigenic sites were labeled on the thin sections of the acrosomal vesicles, by applying the protein A-gold technique. The anti-15.5K lysin antibody was well localized at the posterior half of the acrosomal vesicle.
183S