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61.
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
62.
Salts of transition elements that alter the rate of sperm cell movement act at or near calcium-binding sites. After living bull sperm cells had been preincubated in VO43?, Ni2+, Zn2+, Mn2+, and also La3+, they were then fixed. Crisply defined organelles and the absence of particulate deposits in the morphological controls contrasted sharply with the treated specimens; the latter contained regions of increased electron density, the nature and distribution of which depended on the test substance, reflecting the differential affinities of the specific ions. La3+ formed fine dense areas, mainly at the exocytic surface of the plasma membrane. VO43? marks the cell surface but also left particulate densities within the cell. Ni2+ caused a nearly uniformly dense deposit at the surface and on the satellite fibers and axonemal microtubules. Zn2+ formed less uniform but coarser deposits, while in Mn2+ the distribution was similar to that in Zn2+ but much denser in the axonemal matrix and on the satellite fibers. Verapamil restricted the size and number of the opacities, while procaine permitted a similar distribution of slightly larger size reaction product. The differences in size and distribution of the enhanced densities were consistent and replicable for the individual assay substances. Vanadate, which specifically inhibits Na, K-ATPase, bound to ouabain-sensitive enzyme loci, however, completely disrupting the axonemal complex. This suggests that an important role of dynein in flagellar motion may relate to intracellular transport of Ca2+. 相似文献
63.
Altered growth kinetics precede calcium-induced differentiation in mouse epidermal cells 总被引:1,自引:0,他引:1
K. Elgjo H. Hennings O. P. F. Clausen 《In vitro cellular & developmental biology. Plant》1986,22(6):332-336
Summary Primary cultures of newborn mouse epidermal cells proliferate rapidly and with a high growth fraction for several months when
grown in medium with low calcium (0.02 to 0.1 mM). Addition of calcium to levels generally used in culture medium (1.2 mM) was followed by rapid changes in the pattern of proliferation. By using a combination of technics (a stathmokinetic method,
autoradiography, [3H]thymidine incorporation into DNA, DNA flow cytometry) it was found that cell flux was blocked for 5 to 6 h, followed by
a short rise in the mitotic rate at 10 h, and a gradual fall in all growth parameters until about 32 h after the calcium switch.
There was no accumulation of cells in any particular cell cycle phase. The results indicate that the calcium switch is followed
by a strong reduction in cell flux from G1 whereas the majority of the cells that had left G1 at the time of the switch completed one cell division before cessation of all proliferative activity. Both before and after
the switch the primary epidermal cultures consisted of one diploid and one tetraploid G1 DNA stemline that seemed to react in the same way to calcium.
This work reported in this paper was undertaken during the tenure of an American Cancer Society-Eleanor Roosevelt-International
Cancer Fellowship awarded by the International Union Against Cancer (K. E.). The project was supported by funds partly provided
by the International Cancer Research Data Bank Program of the National Cancer Institute, National Institutes of Health, Bethesda,
MD, under contract N01-C0-65341 (International Cancer Research Technology Transfer) and partly by the International Union
Against Cancer (O.P.F.C.). 相似文献
64.
Summary In the isolated bullfrog cornea, three calcium channel antagonists had dose-dependent inhibitory effects on the Cl-originated short-circuit current (SCC). Their order of decreasing potency was bepridil, verapamil and diltiazem. One millimolar diltiazem inhibited the SCC by 98% and subsequent incubation with the calcium ionophore A23187 had no restorative effect. Increasing the bathing solution Ca concentration from 0.05 to 15mm, however, decreased diltiazem's inhibitory efficacy. This antagonist depolarized the intracellular potential differenceV
m
from –54 to –18 mV (tear: reference) and the voltage divider ratioFR
0 decreased from 0.58 to 0.30, suggesting an increase in basolateral membrane electrical resistance. Additional indication of a basolateral membrane effect by the drug was that preincubation with 105
m amphotericin B in Cl-free Ringer's did not eliminate the inhibitory effect of the drug on the Na- and K-elicited SCC. In the absence of amphotericin B in Cl-free Ringer's (SCC=0), 1 ×103
m diltiazem depolarized theV
m
from –78 to –9 mV suggesting that the increase in basolateral membrane resistance was due to K channel blockade. Diltiazem (1×103
m) significantly decreased cyclic AMP content; however, isoproterenol in the presence of the drug increased cyclic AMP fourfold without having any restorative effect on the inhibited SCC. Therefore, the inhibition of the Cl-originated SCC resulting from an increase in basolateral membrane K resistance is not caused by a decline in cyclic AMP content. In plasma membrane-enriched fractions prepared from broken cell preparations of bovine corneal epithelium, 1×103
m diltiazem had no inhibitory effects on either Na,K-ATPase or Ca,Mg-ATPase activities. These latter effects further point to the selectivity of diltiazem as an inhibitor of K-channel activity, but do not preclude a Ca-channel blocker effect by the drug in the micromolar range. 相似文献
65.
66.
ZINC TOLERANCE IN BETULA 总被引:4,自引:2,他引:2
67.
Solanum elaeagnifolium Cav. fruits contain high concentrations of steroidal saponins. Treatment of 3-day-old clover seedlings with aqueous fruit extracts modified Ca2+ uptake without significantly altering K+ and H2 PO4 − uptake. The extracts increased Ca2+ uptake in the concentration range of 0.2 to 20 m M Ca2+ . Uptake curves could be represented by two phases. In the lower phase (0.2-1.0 m M Ca2+ ), this change could be related to an increase in Vmax . Pretreatment of seedlings with saponin extracts significantly reduced ATP-dependent Ca2+ uptake and Ca2+ -dependent ATPase activity in a fraction isolated from root homogenates by centrifugation at 1500 g for 15 min. Saponins purified from S. eleagnifolium extracts by thin-layer chromatography modified in vitro the Ca2+ -ATPase activity of this fraction, indicating that the steroid may act directly on Ca2+ transport across membranes. 相似文献
68.
The effects of intracerebroventricular (i.c.v.) administration of calcitonin and PGE2 on intestinal motility and body temperature were examined in conscious rats chronically fitted with intraparietal electrodes in the small intestine, a cannula in a cerebral lateral ventricle and a subcutaneous thermistor probe. Both calcitonin and PGE2 restored the fasted pattern of intestinal motility in fed rats and induced an increase in body temperature. Indomethacin, an inhibitor of the cyclooxygenase with calcium antagonistic properties, and TMB-8, an intracellular calcium antagonist, blocked the effects of calcitonin on intestinal motility and body temperature. Piroxicam, an inhibitor of the cyclooxygenase which does not affect calcium uptake blocked the thermic but not the intestinal effects of calcitonin. TMB-8 but not indomethacin or piroxicam partially blocked the effects of PGE2 on both intestinal motility and body temperature. It is concluded that the central hyperthermic effect of calcitonin is mediated through the formation and the release of prostaglandins whereas the central action of calcitonin on digestive motility results from intracerebral effects on calcium fluxes. 相似文献
69.
Computer search of calcium binding sites in a gene data bank: use of learning techniques to build an expert system 总被引:3,自引:0,他引:3
Using a learning set of 28 sequences able to bind calcium (each sequence is 12 residues long), we have built two filters by learning on this set. The first filter uses a pattern-matching technique and the second one takes into account the environment of amino-acids. These two filters have been used to find new calcium-binding proteins in a data bank. The results are discussed. 相似文献
70.
To improve the inulinase (2,1-β-d-fructan fructanohydrolase, EC 3.2.1.7) stability of calcium alginate-immobilized Kluyveromyces marxianus cells, treatment with hardening agents has been investigated. Treatment of immobilized cells with some polycationic polymers resulted in little decrease in volumetric reactor productivity, but was most effective in increasing the inulinase stability of the immobilized cells. Inulinase stability of glutaraldehyde-hardened immobilized cells increased two-fold, and for hexamethylenediamine + glutaraldehyde and polyethyleneimine + glutaraldehyde-hardened cells increased six-fold compared with that of the unhardened cells. 相似文献