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1.
Spermatozoa of the sea urchin, Hemicentrotus pulcherrimus (108 cells/ml), preincubated with unfertilized eggs deprived of jelly coats (more than l05 cells/ml) at 20°C for 20min in Mg2+ free artificial sea water containing 1 mM Ca2+ (MFASW), exhibited very low respiration, which was enhanced by 2, 4 dinitrophenol (DNP). The fertilization rate in MFASW was usually less than 5% and was about 25% at most. Preincubation with fertilized eggs (with and without a fertilization membrane) in MFASW did not reduced the respiratory rate of spermatozoa. The rate of sperm respiration was lower in MFASW than in artificial sea water (ASW), but was higher than the respiratory rate of spermatozoa preincubated in MFASW with unfertilized eggs. Sperm respiration in MFASW or in ASW was not stimulated by 2, 4 dinitrophenol. Almost complete inhibition of sperm respiration was obtained with unfertilized eggs fixed with glutaraldehyde at concentrations of above 105 cells/ml in MFASW and of about l04 cells/ml in ASW. The respiratory rate of spermatozoa treated with fixed eggs was enhanced by DNP. It is concluded that the respiratory rate of the spermatozoa is reduced by their interaction with unfertilized eggs before their penetration into the eggs. 相似文献
2.
AKIKO FUJIWARA AKIYA HINO TAKAAKI HIRUMA IKUO YASUMASU 《Development, growth & differentiation》1982,24(2):145-154
In spermatozoa of all examined sea urchins, the respiration was inhibited and their motility was lowered by the glutaraldehyde-fixed eggs. The respiration of the fixed-egg-reacted spermatozoa was stimulated by 2, 4 dinitrophenol in Clypeaster japonicus, Hemicentrotus pulcherrimus and pseudocentrotus depressus but was not in Anthocidaris crassispina and Toxopneustes pileolus. Ratio of ADP to ATP was markedly lower in the reacted spermatozoa of the former species than in those of Anthocidaris. The low respiratory rate in the former species probably results from ADP control but does not in the latter species. Tetramethyl-p-phenylenediamine enhanced the respiratory rate in the reacted spermatozoa of the latter species to almost the same rate as in the intact spermatozoa, but elevated slightly in the former species. The inhibition of electron transport in mitochondrial respiratory chain is probably predominant in the latter species. In the former species, the slight inhibition of electron transport does not seem to result in a failure of ADP phosphorylation, and hence the stop of movement probably causes a shortage of ADP. Carnitine, which made the reacted spermatozoa of all species motile, enhanced the respiratory rate only in those of the former species. 相似文献
3.
Eggs of the sea urchin, Hemicentrotus pulcherrimus , were stimulated by halothane, known to induce Ca2+ release from sarcosome, to cause fertilization membrane formation in normal and Ca2+ free artificial sea water. In the absence of external Ca2+ , halothane-induced formation of fertilization membrane was inhibited by dantrolene, an inhibitor of Ca2+ release from sarcosome, but was not blocked by nifedipine, a Ca2+ antagonist specific to Ca2+ channels in plasma membrane. Ca2+ release from sedimentable fraction isolated from eggs was induced by halothane and was inhibited by dantrolene, but was not blocked by nifedipine. In normal artificial sea water, halothane-caused egg activation was not inhibited either by dantrolene or by nifedipine, but was blocked in the presence of both compounds. 45 Ca2+ influx was substantially stimulated by halothane in eggs exposed to 45 CaCl2 . Halothane-induced 45 Ca2+ influx into eggs was inhibited by nifedipine but was not blocked by dantrolene. When Ca2+ release from intracellular organellae is blocked, Ca2+ transport through Ca2+ channels in plasma membrane probably acts as a "fail-safe" system to induce an increase in cytosolic Ca2+ level, resulting in egg activation. 相似文献
4.
When sperm of the sea urchin, Hemicentrotus pulcherrimus, are diluted into K+ -free seawater, the pH of the suspension gradually decreases, whereas a rapid decline in pH is observed following dilution into regular seawater. Sperm motility and respiration are also activated after dilution into K+ -free seawater, but levels of activity are less than those observed following dilution into regular seawater. Upon addition of 10 mM K+ to K+ -free seawater, rapid acid release occurs and motility and respiratory rate in sperm are reactivated. The effect of K+ on respiration was competitive with respect to the external Na+ concentrations. Harmaline, a potent inhibitor of Na+ /K+ -ATPase, causes a decrease in movement and respiration of the sperm. Harmaline does not inhibit the rapid decline in pH, although it depresses the release of acid from mitochondria. These results suggest that external K+ plays an important role in intracellular alkalinization of sea urchin sperm. 相似文献
5.
AKIKO FUJIWARA MASATOSHI MITA AKIYA HINO TOSHIKAZU HAMASAKI YUTAKA NAITOH IKUO YASUMASU 《Development, growth & differentiation》1983,25(1):39-47
Spermatozoa of the sea urchin, Hemicentrotus pulcherrimus , showed marked decrease in respiration, and arrested movement after interaction with the fixed eggs. Immotile spermatozoa that had reacted with fixed eggs contained higher levels of long chain fatty acyl-CoAs than normal motile spermatozoa. On treatment with carnitine, the immotile spermatozoa became motile again and their intracellular concentrations of long chain fatty acyl-CoAs decreased. On incubation with anti-mycin A or CN− for 20 min, the motility of normal spermatozoa decreased gradually but their long chain fatty acyl-CoA content changed only slightly. The decrease in sperm motility in the latter case was probably due to decrease in the level of ATP, resulting from inhibition of respiration by antimycin A or CN− . The motility of spermatozoa extracted with Triton X-100 was restored by ATP and their movement was inhibited by long chain fatty acyl-CoAs, such as myristoly CoA and palmitoyl-CoA, but was not by short chain fatty acyl-CoAs, such as acetyl-CoA, propionyl CoA and butyryl-CoA. Na-palmitate, Na-myristate and CoA did not inhibit the reactivation of extracted spermatozoa by ATP. 相似文献
6.
In unfertilized eggs of the sea urchin Hemicentrotus pulcherrimus , fertilization membrane formation was induced by an incubation with dimethylsulfoxide (DMSO) for several min at 20°c followed by another incubation in an ice bath. The number of eggs with fertilization membrane, thus obtained, increased in relation to the concentration of DMSO between 1 and 3% (v/v) and was higher than 75% at concentrations above 3%. Fertilization membrane formation by this treatment occurred in Ca2+ free- or Ca2+ , Mg2+ free- artificial sea water containing EGTA (50 mM) and was inhibited by verapamil. In the presence of DMSO, the membrane formation was also induced by 2, 4-dinitrophenol or cyanide in considerable number of eggs at 20°c. Eggs remained fertilizable, even when they were kept with DMSO for 1 hr at 20°c. DMSO slightly enhanced respiratory rate in unfertilized eggs and substantially reduced it in fertilized eggs. DMSO-treated eggs exhibited cyanide-insensitive respiratory burst following chilling in an ice bath or by adding DNP or cyanide, in a similar manner to the burst induced by sperm. 相似文献
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8.
Abstract Bacteroides fragilis Bf-2 cells were more sensitive to far-UV radiation, N -methyl- N '-nitrosoguanidine, ethylmethane sulphonate, acriflavine and mitomycin C under aerobic conditions than under anaerobic conditions. The opposite effect was observed with H2 O2 -treated cells and exposure to O2 enhanced the survival of H2 O2 -treated cells. Pretreatment of cells with sublethal concentrations of H2 O2 also increased the survival of H2 O2 -treated cells. Reactivation of UV- and X-irradiated and methylmethane sulphonate and H2 O2 -treated phage b-1 was induced by O2 and H2 O2 in B. fragilis . 相似文献
9.
Kenneth Hensley Quentin N. Pye Michael L. Maidt Charles A. Stewart Kent A. Robinson Fatima Jaffrey Robert A. Floyd 《Journal of neurochemistry》1998,71(6):2549-2557
Abstract: Mitochondrial complexes I, II, and III were studied in isolated brain mitochondrial preparations with the goal of determining their relative abilities to reduce O2 to hydrogen peroxide (H2 O2 ) or to reduce the alternative electron acceptors nitroblue tetrazolium (NBT) and diphenyliodonium (DPI). Complex I and II stimulation caused H2 O2 formation and reduced NBT and DPI as indicated by dichlorodihydrofluorescein oxidation, nitroformazan precipitation, and DPI-mediated enzyme inactivation. The O2 consumption rate was more rapid under complex II (succinate) stimulation than under complex I (NADH) stimulation. In contrast, H2 O2 generation and NBT and DPI reduction kinetics were favored by NADH addition but were virtually unobservable during succinate-linked respiration. NADH oxidation was strongly suppressed by rotenone, but NADH-coupled H2 O2 flux was accelerated by rotenone. α-Phenyl- N-tert -butyl nitrone (PBN), a compound documented to inhibit oxidative stress in models of stroke, sepsis, and parkinsonism, partially inhibited complex I-stimulated H2 O2 flux and NBT reduction and also protected complex I from DPI-mediated inactivation while trapping the phenyl radical product of DPI reduction. The results suggest that complex I may be the principal source of brain mitochondrial H2 O2 synthesis, possessing an "electron leak" site upstream from the rotenone binding site (i.e., on the NADH side of the enzyme). The inhibition of H2 O2 production by PBN suggests a novel explanation for the broad-spectrum antioxidant and antiinflammatory activity of this nitrone spin trap. 相似文献
10.
Katsuaki Hoshino Takeshi Shimizu Yutaka Sendai Tatsuo Harumi Norio Suzuki 《Development, growth & differentiation》1992,34(4):403-411
We examined the effects of two egg jelly components, a fucose sulfate glycoconjugate (FSG) and sperm-activating peptide I (SAP-I: Gly-Phe-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly), on the intracellular pH (pHi ) and Ca2+ ([Ca2+ ]i ) of spermatozoa of the sea urchin Hemicentrotus pulcherrimus . FSG and/or SAP-I induced elevations of [Ca2+ ]; and pHi in the spermatozoa at pH 8.0. At pH 8.0, a second addition of FSG did not induced further elevation of the [Ca2+ ]i or pHi of spermatozoa treated with FSG, but addition or FSG after SAP-I or of SAP-I after FSG induced further increases of [Ca2+ ]i and pHi , At pH 6.6, FSG and/or SAP-I did not induce significant elevation of the [Ca2+ ]i , although SAP-I elevated the pHi , its half-maximal effective concentration being 10 to 100 pM. At pH 8.0, tetraethyl-ammonium, a voltage-sensitive K+ -channel blocker, inhibited induction of the acrosome reaction and elevations of [Ca2+ ]i and pHi by FSG, but did not affect those by SAP-I. These results suggest that FSG and SAP-I activate different Ca2+ and H+ transport systems. 相似文献
11.
The Dutch elm disease (DED) pathogen Ophiostoma novo-ulmi Buissm. elicited the production of H2 O2 in cell suspension cultures of the resistant species Ulmus pumila L. This response was not observed in suspensions of the susceptible elm U. campestris Mill. H2 O2 production started after a lag time of 30–40 min following inoculation, peaked between 4 and 6 h and lasted up to 24 h. Treatment of the suspensions with exogenously added H2 O2 did not cause accumulation of the sesquiterpene phytoalexins mansonones nor of the coumarin scopoletin. Spore germination and growth of O. novo-ulmi were significantly delayed with different amounts of H2 O2 (0.1–1 m M ). These results suggest that H2 O2 production is an inducible defence response which may contribute to DED resistance by delaying the growth of the pathogen at the earliest stages of infection. Whether H2 O2 is involved in other elm defence responses to the pathogen is presently unknown, but its production seems to be an independent event from phytoalexin formation. 相似文献
12.
Inhibition of Dopamine Release by Prostaglandin EP3 Receptor via Pertussis Toxin-Sensitive and -Insensitive Pathways in PC12 Cells 总被引:1,自引:0,他引:1
Kazuhiro Nakamura Hironori Katoh Atsushi Ichikawa Manabu Negishi 《Journal of neurochemistry》1998,71(2):646-652
Abstract: Hydrogen peroxide (H2 O2 ) is produced from several sources in brain and may be involved in neurodegeneration and second messenger signaling. Little is known about the effects of H2 O2 on transmitter storage in brain synaptic vesicles. Neurotransmitter uptake into synaptic vesicles is driven by an electrochemical proton gradient generated by the vacuolar H+ -ATPase (V-ATPase) in the vesicle membrane. We report here that the V-ATPase in bovine brain synaptic vesicles is highly sensitive to inhibition by micromolar concentrations of H2 O2 . Glutamate uptake by the vesicles is also inhibited, very likely as a secondary consequence of ATPase inactivation. Dithiothreitol or reduced glutathione reverse H2 O2 -induced inhibition of the V-ATPase, and ATP or GTP partially protect the ATPase from inhibition by H2 O2 . These and other results suggest that the mechanism of inhibition of the V-ATPase by H2 O2 involves oxidation of a reactive cysteine sulfhydryl group in the ATP binding site. Inhibition of V-ATPase activity would decrease the amount of transmitter stored in synaptic vesicles and thus down-regulate transmitter release during episodes of oxidative stress or in response to second messenger signaling. 相似文献
13.
Abstract: We studied the action of H2 O2 on the exocytosis of glutamate by cerebrocortical synaptosomes. The treatment of synaptosomes with H2 O2 (50–150 µ M ) for a few minutes results in a long-lasting depression of the Ca2+ -dependent exocytosis of glutamate, induced by KCl or by the K+ -channel inhibitor 4-aminopyridine. The energy state of synaptosomes, as judged by the level of phosphocreatine and the ATP/ADP ratio, was not affected by H2 O2 , although a transient decrease was observed after the treatment. H2 O2 did not promote peroxidation, as judged by the formation of malondialdehyde. In indo-1-loaded synaptosomes, the treatment with H2 O2 did not modify significantly the KCl-induced increase of [Ca2+ ]i . H2 O2 inhibited exocytosis also when the latter was induced by increasing [Ca2+ ]i with the Ca2+ ionophore ionomycin. The effects of H2 O2 were unchanged in the presence of superoxide dismutase and the presence of the Fe3+ chelator deferoxamine. These results appear to indicate that H2 O2 , apparently without damaging the synaptosomes, induces a long-lasting inhibition of the exocytosis of glutamate by acting directly on the exocytotic process. 相似文献
14.
Massimo Malerba Paolo Crosti Raffaella Cerana Renato Bianchetti 《Physiologia plantarum》2003,119(4):480-488
Fusicoccin (FC) is a well known toxin acting as a 14-3-3 protein-mediated activator of the plasma membrane H+ -ATPase and it has been widely used to study the regulatory mechanism and the physiological role of this enzyme's activity. Recently, FC has been shown to induce other responses similar to those occurring under a stress condition, perhaps not strictly dependent on the activation of proton extrusion. In this paper we report that in cultured sycamore ( Acer pseudoplatanus L.) cells FC induces H2 O2 overproduction as well as other novel, presumably related responses, such as the activation of the alternative oxidase and the leakage of cytochrome c from the mitochondria, accompanied by a decrease of the cytochrome pathway capacity. The relationship between H2 O2 production and other phenomena has also been studied by means of exogenously added H2 O2 . 相似文献
15.
Ryanodine Activates Sea Urchin Eggs 总被引:3,自引:2,他引:1
C. Sardet I. Gillot A. Ruscher P. Payan J.-P. Girard G. de Renzis 《Development, growth & differentiation》1992,34(1):37-42
We have studied the effect on sea urchin eggs of ryanodine, a plant alkaloid that causes muscle contraction by opening calcium channels in the sarcoplasmic reticulum terminal cisternae. Ryanodine, although it is less effective that IP3 , produces full or partial activation in 62% of injected sea urchin eggs. In addition ryanodine inhibits in a dose dependant manner 45 Ca pumping in the isolated egg cortex or in eggs permeabilized with digitonin. Efflux experiments show that in fact ryanodine as IP3 stimulates the release of calcium sequestered intracellularly. We further show that these effects of ryanodine are inhibited by Mg++ , ruthenium red and heparin. Our results suggest that ryanodine-sensitive intracellular calcium channels exist in the sea urchin egg. 相似文献
16.
The role of the ascorbate-glutathione cycle and AOS detoxification was investigated during leaf growth of defoliated and undefoliated plants of ryegrass ( Lolium perenne L. cv. Bravo). Antioxidants and related enzymatic activities were located in elongating leaf bases (ELBs) of undefoliated plants, following a decreasing gradient from basal (meristem) to distal segments, inverse to H2 O2 levels. In the meristematic zone, the intense activity of the ascorbate-glutathione cycle and the supply of reducing power by the oxidative pentose phosphate pathway allowed the maintenance of both antioxidant reduction and H2 O2 detoxification. BCNU (1–3 bis(2-chloroethyl)- N -nitrosourea), a glutathione reductase inhibitor, induced an increase in the meristematic zone in both H2 O2 and antioxidant levels and a decrease in reduced/oxidized ratios of glutathione and ascorbate. These changes were associated with a reduced foliar regrowth activity. In the absence of BCNU, defoliation did not modify the ratios of reduced/oxidized antioxidants, although it triggered a temporary increase in H2 O2 level. The results are discussed on the basis of a possible control of leaf growth by glutathione and ascorbate. 相似文献
17.
Tatsuo Harumi Katsuaki Hoshino Norio Suzuki 《Development, growth & differentiation》1992,34(2):163-172
The effects of sperm-activating peptide I (SAP-I: Gly-Phe-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly) on Hemicentrotus pulcherrimus spermatozoa in high [K+ ] sea water were examined. In high [K+ ] sea water, the respiration rates and motility of H. pulcherrimus spermatozoa were lower than those in normal sea water. SAP-I did not stimulate the lowered respiration rate or motility, although the peptide bound to the spermatozoa as it does in normal sea water. SAP-I elevated the sperm cGMP level in 100 mM K+ sea water (from 0.37 to 4.81 pmol/mg wet weight spermatozoa) more than those in normal sea water (from 0.21 to 0.93 pmol/mg wet weight). A phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX) and SAP-I synergistically elevated the cGMP level from 0.35 to 33.08 pmol/mg wet weight in 100 mM K+ sea water. However, in high [K+ ] sea water, SAP-I did not increase the cAMP level even in the presence of IBMX. SAP-I caused rapid, transient elevation of the intracellular pH and Ca2+ concentration of spermatozoa in normal sea water but not in 100mM K+ sea water. SAP-I did not decrease the apparent molecular weight of sperm guanylate cyclase from 131,000 to 128,000 in high [K+ ] sea water. These results suggest that the SAP-I-induced elevation of the cGMP level in sea urchin spermatozoa occurs before or independently of membrane hyperpolarization induced by the opening of K+ channels. 相似文献
18.
Tatsuo Harumi Masaaki Yamaguchi Norio Suzuki 《Development, growth & differentiation》1991,33(1):67-73
Sperm-activating peptide analogues, GYGG-SAP-I (Gly-Tyr-Gly-Gly-Gly-Phe-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly) and GYGG-SAP-IIB (Gly-Tyr-Gly-Gly-Lys-Leu-Cys-Pro-Gly-Gly-Asn-Cys-Val) which exibit almost the same respiration-stimulating activity as the respective original peptide were chemically synthesized, radiolabeled with Na125 1, and used for experiments of binding and cross-linking to Hemicentrotus pulcherrimus or Glyptocidaris crenularis spermatozoa. In competitive binding, SAP-I caused 50% decrease in 125 l-GYGG-SAP-l binding to intact spermatozoa, sperm heads and sperm tails of H. pulcherrimus at the concentrations of 282 nM, 3 nM and 141 nM, respectively. The incubations of H. pulcherrimus sperm tails with 125 l-GYGG-SAP-I and the chemical cross-linking reagent, disuccinimidyl suberate, resulted in the appearance of two radiolabeled bands with apparent molecular masses of 71 kDa and 63 kDa (determined by sodium dodecyl sulfate-gel electrophoresis under reducing conditions). 125 l-GYGG-SAP-IIB bound almost equally to sperm heads and sperm tails of G. crenularis , and was cross-linked to 172 kDa, 62 kDa and 58 kDa proteins in sperm heads, and 157 kDa and 62 kDa proteins in sperm tails with disuccinimidyl suberate. 相似文献
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Roles of H2 O2 in the infection process of Magnaporthe oryzae on rice were investigated. In a leaf sheath assay for up to 48 h post-inoculation, the absence or presence of catalase in the conidia suspension was correlated with the level of accumulated H2 O2 in infected leaf cells, as observed by staining with 3',3-diaminobenzidine tetrahydrochloride. In the incompatible interaction, the appearance of autofluorescence or frequency of cell death characterized by granulation (symptoms characteristic of hypersensitive responses) was not significantly affected by the presence of catalase in the conidia suspension. In the leaf blade assay, inoculation of compatible conidia in the presence of catalase produced more severe symptoms than that of conidia in the absence of catalase at 6 days post-inoculation. These results suggest that, in this host–parasite interaction, the primary role of host-produced H2 O2 is in limiting hyphal growth after penetration through toxic action. Furthermore, in incompatible interactions, H2 O2 is implied not to be a major mediator of hypersensitive cell death. 相似文献