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31.
E. R. S. Roldan S. Shibata R. Yanagimachi 《Molecular reproduction and development》1986,13(4):281-292
The effects of Ca2+ channel antagonists on the motility and acrosome reaction of guinea pig spermatozoa were examined by incubating the spermatozoa continuously in Ca2+-containing capacitating media with 10?6 M to 10?4 M antagonist. Antagonists tested were four voltage-gated Ca2+ channel antagonists (verapamil, nifedipine, nimodipine, and FR–34235) and two ligand-gated channel antagonists (NaNO2 and Na-nitroprusside). None of these antagonists could block the acrosome reaction. Instead, three antagonists (verapamil, nimodipine, and FR-34235, each at 10?4 M) accelerated the onset of the acrosome reaction with a subsequent decrease in sperm motility. Nifedipine and Na-nitroprusside at the same concentration caused a complete loss of sperm motility by 4 hr of incubation with no substantial effect on the rate of acrosome reaction. The detrimental effect of antagonists on the motility of spermatozoa appears to be due to a direct, Ca2+-independent, membrane-perturbing action of the reagents. The acrosome reaction was not inhibited when guinea pig spermatozoa were precapacitated in Ca2+-free medium (with a low concentration of lysolecithin) in the continuous presence of antagonists. An acceleration of the onset of the acrosome reaction by verapamil (10?4 M) was also demonstrated in the golden hamster. These results may be interpreted as indicating that the entry of extracellular Ca2+ into spermatozoa, which triggers the acrosome reaction of guinea pig and hamster spermatozoa, is not mediated by Ca2+ channels. This is in marked contrast with the case reported in invertebrate spermatozoa. Possible mechanisms by which some of the antagonists stimulate the acrosome reaction and affect the motility of mammalian spermatozoa are discussed. 相似文献
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Adsorption of chloroform, halothane, enflurane and diethyl ether on the air/water interface was compared with adsorption on the dipalmitoylphosphatidylcholine monolayer, spread on the air/water interface, at four compressional states; 88.5, 77.0, 66.5 and 50.5 A2 surface area per phosphatidylcholine molecule. Anesthetics were administered from the gas phase. The affinities of these agents to the phosphatidylcholine monolayer varied according to the state of the monolayer. Chloroform and halothane showed a stronger affinity to the highly compressed phosphatidylcholine monolayer (50.5 A2) than to the expanded monolayer (88.5 A2) or to the air/water interface without the monolayer. Diethyl ether behaved in reverse; a stronger affinity to the expanded monolayer was exhibited than to the compressed monolayer. Enflurane showed the highest affinity to the intermediately compressed monolayer (77.0 A2). The adsorption isotherm of anesthetics to the monolayer was characterized by atypical Langmuir-type, in which available number of binding sites changed when anesthetics were adsorbed. The mode of adsorption onto the monolayer was dissimilar to adsorption onto air/water interface, where adsorption followed the Gibbs surface excess. A theory is presented to explain the above differences. The adsorbed anesthetic molecules do not stick to phosphatidylcholine molecules but penetrate into the monolayer lattice and occupy the phosphatidylcholine sites at the interface. Quantitative agreement between the theory and the experimental data was excellent. For the monolayer at 50.5 A2 compression, the changes in the transfer free energy accompanying the anesthetic adsorption from the gas phase to the monolayer were in the order of chloroform greater than halothane greater than enflurane greater than diethyl ether, in agreement with the clinical potencies. 相似文献
35.
Polymorphism of soluble aconitase was investigated in 152 Japanese placentae. The allelic frequencies were ACONS1 = 0.951 and ACONS2 = 0.049. ACONS2 appears to be rather high among Orientals including Japanese, while ACONS4 seems to be characteristic for Negroids. 相似文献
36.
To find whether cytoplasmic streaming in Acetabularia is controlledby Ca2+, a tonoplast-permeabilized cell model was prepared usinga vacuolar perfusion technique. The cytoplasmic streaming remainedalmost normal after perfusion with EGTA medium (10 mM EGTA,40 mM PIPES, 5mM MgCl2 and 800 mM sorbitol, pH 6.9), but stoppedwithin 10 min when saponin medium (EGTA medium plus 50 µg/mlsaponin, 50 µg/ml hexokinase and 5 mM glucose) was perfused.This model system was reactivated with a solution containing0.5 mM ATP and different concentrations of Ca2+ (reactivationmedium). With the reactivation medium at pCa 65, theresumed streaming lasted for about 10 min before the cytoplasmaggregated. At pCa 43, the streaming was observed onlyfor a few minutes because the cytoplasm aggregated quickly.At pCa 7, no reactivated movement was observed. Reactivationwas not induced in an ATP- or Mg2+-deficient medium even inthe presence of an adequate concentration of Ca2+, and was inhibitedby 50 µg/ml cytochalasin B or 1 mM N-ethylmaleimide. We concluded from these observations that the cytoplasmic streamingin Acetabularia is very likely to be driven by the actomyosinsystem in the presence of Mg-ATP and Ca2+ at pCa 65. (Received October 31, 1984; Accepted April 1, 1985) 相似文献
37.
Purification, characterization, and immunological properties of fumarase from Euglena gracilis var. bacillaris. 总被引:3,自引:1,他引:2
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A rapid three-step procedure utilizing heat treatment, ammonium sulfate fractionation, and affinity chromatography on Matrex gel Orange A purified fumarase (EC 4.2.1.2) 632-fold with an 18% yield from crude extracts of Euglena gracilis var. bacillaris. The apparent molecular weight of the native enzyme was 120,000 as determined by gel filtration on Sephacryl S-300. The preparation was over 95% pure, and the subunit molecular weight was 60,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that the enzyme is a dimer composed of two identical subunits. The pH optimum for E. gracilis fumarase was 8.4. The Km values for malate and fumarate were 1.4 and 0.031 mM, respectively. Preparative two-dimensional gel electrophoresis was used to further purify the enzyme for antibody production. On Ouchterlony double-immunodiffusion gels, the antifumarase serum gave a sharp precipitin line against total E. gracilis protein and purified E. gracilis fumarase. It did not cross-react with purified pig heart fumarase. On immunoblots of purified E. gracilis fumarase and crude cell extracts of E. gracilis, the antibody recognized a single polypeptide with a molecular weight of approximately 60,000, indicating that the antibody is monospecific. This polypeptide was found in E. gracilis mitochondria. The antibody cross-reacted with an Escherichia coli protein whose molecular weight was approximately 60,000, the reported molecular weight of the fumA gene product of E. coli, but it failed to cross-react with proteins found in crude mouse cell extracts, Bacillus subtilis extracts, or purified pig heart fumarase. 相似文献
38.
S Ichikawa T Shibata Y Takehara H Tamada K Oda S Murao 《Journal of reproduction and fertility》1985,73(2):385-390
Proteinase inhibitors of microbial origin were injected into the uterine horns of mated rats at 14:00 h on Day 5 of pregnancy (spermatozoa in vaginal smear = Day 1), and 5 or 6 h later the embryos were flushed from the horns and examined. Chymostatin and alpha-MAPI, inhibitors of chymotrypsin-like serine proteinase and thiol proteinases, as well as thiolstatin, an inhibitor of thiol proteinases, significantly inhibited embryo growth. The inhibitory activity of alpha-MAPI on embryonic growth was distinctly greater than that of thiolstatin, although the ID50 values of the two inhibitors to papain are similar. Antipain and leupeptin which are inhibitors of trypsin-like and thiol proteinases, and talopeptin, an inhibitor of metal proteinases, significantly interrupted the removal of the zona pellucida from expanding blastocysts. These results suggest that (1) a chymotrypsin-like proteinase seems to be important to the growth of the embryo, (2) a thiol proteinase may participate in embryonic growth, and (3) a trypsin-like proteinase and a metal proteinase are likely to participate in zonalysis. 相似文献
39.
Makoto Daimon Ken-ichi Tsutsumi Jun-ichi Sato Reiko Tsutsumi Kiichi Ishikawa 《Biochemical and biophysical research communications》1984,124(2):337-343
The expression of aldolase A and B mRNAs during azo-dye-induced carcinogenesis in rat liver was examined. After feeding the dye for 18 weeks, the level of aldolase A mRNA increased to about 11 times that in a normal liver, with the concomitant decrease of aldolase B mRNA level to about 25% of that in a normal liver. These changes did not occur progressively during the carcinogenesis, but occurred as an additional phase after 4 week-feeding of the azo-dye. At this stage, the levels of aldolase A and B mRNAs were about 7 times and 45% of that in a normal liver, respectively. This biphasic pattern in the aldolase isozyme expression in the azo-dye-fed rat liver is discussed together with the kinetic data of the enzyme activity. 相似文献
40.
Summary The vacuolar system in the absorptive cells of the goldfish hindgut was studied by rapid freeze-substituted and cytochemical techniques. The apical cytoplasm of the absorptive cells contained two types of vacuoles: endosomes and lysosomes. The former were characterized by an absence of acid phosphatase activity, a dot-like distribution of material at the peripheral rim, the labelling of the inner surface with horseradish peroxidase (HRP), and by frequent connections to cytoplasmic tubules (CT), which were also free of acid phosphatase activity. The latter vacuole was preferentially located in the deeper cytoplasm and was characterized by the presence of acid phosphatase activity, an electron-dense interior matrix, a peripheral electron-lucent region (a halo), and by the detachment of HRP from the inner surface. Connections between CTs and these latter vacuoles were rarely seen. In the deeper cytoplasm, fusion between endosomes and lysosomes was sometimes observed. These results suggest that the vacuoles which are associated with CTs are endosomes, but not lysosomes, and that internalized materials are transported through the endosome-lysosome system to a giant food vacuole in the cell. 相似文献