Sperm chymotrypsin-like enzymes of different inhibitor-susceptibility as lysins in ascidians

Inhibitory effects of three peptidyl phenylalaninals on fertilization and on chymotrypsin-like enzyme activity of sperm in three species of ascidians were examined. The results suggest that a sperm chymotrypsin-like enzyme is indispensable for the fertilization in each of the ascidians, and that these enzymes have different susceptibilities to inhibitors.     

Timing of action of sperm proteases in ascidian fertilization

Summary The timing of action of three sperm proteases, acrosin, spermosin, and a chymotrypsin-like enzyme, in the fertilization of the ascidian,Halocynthia roretzi, was examined by adding specific protease inhibitors at various times after insemination. The results indicate that the last two enzymes both function at the early stage of the process of sperm penetration through the egg investment, while acrosin functions at the late stage.Acknowledgments. We are indebted to Dr M. Hoshi of Tokyo Institute of Technology for his helpful discussion, and to Dr T. Someno of Nippon Kayaku Kogyo Co. for his generous gifts of Z-Val-Pro-Arg-H and leupeptin. This work was supported by Grants-in-Aid from the Ministry of Education, Science and Culture of Japan, and from Naito Research Foundation.     

Involvement of prolyl endopeptidase in ascidian fertilization

Summary Inhibitory effects on fertilization of the ascidian of three benzyloxycarbonyl(Z)-aminoacyl prolinals and Z-Gly-Pro-chloromethyl ketone added before and after insemination were examined. The results suggest that the prolyl endopeptidase is involved in the process of fertilization, especially in a process taking place between chorion elevation and cell cleavage.Acknowledgment. We are indebted to Dr T. Numakunai of the Marine Biological Station of Tohoku University for collecting the ascidians and to Dr M. Hoshi of the Tokyo Institute of Technology for his helpful discussion. We are also grateful to Dr T. Yoshimoto and Prof. D. Tsuru of Nagasaki University for their generous gift of Z-Gly-Prochloromethyl ketone. This work was supported by a Grant-in-Aid from the Ministry of Education, Science and Culture of Japan.     

Lithium and phorbol ester modify the activating capacity of ascidian spermatozoa

In this paper we have shown, using the whole-cell voltage clamp technique, that two parameters of the fertilization current in ascidian eggs may be modified by exposing spermatozoa to lithium or to phorbol ester. When spermatozoa were pre-treated in 250 mM lithium sea water for up to 30 min there was a significant increase in the mean initial slope of the fertilization current, from 116±90 to 169±84 pA/s (p<0.05). The peak current increased from 1371±1079 to 1719±1052 pA (p>0.05). Pre-treatment in 200–600 nM phorbol 12-myristate 13-acetate also increased the activating capacity of ascidian sperm, as monitored by a significant increase in the initial slope current in control eggs; however, there was no increase in peak current. Furthermore, we have shown, using NH₄Cl, that an increase in intracellular pH alone is insufficient to change the activating capacity of spermatozoa. This is the first report showing that the kinetics of an egg activation event depend upon the physiological status of the spermatozoon.     

Vanadium biochemistry: The unknown role of vanadium-containing cells in ascidians (sea squirts)

Summary This article reviews several new developments in vanadium biochemistry, as elucidated from studies of ascidians. A hypothesis correlating ascidian blood cell function to anaerobiosis, via two prominent redox constituents, namely vanadium(III) and the tunichromes, a family of metal ion complexing/reducing hydroquinonoid peptides, is presented.     

Effect of urochordamines on larval metamorphosis of ascidians

Two pteridine-containing bromophysostigmine alkaloids, urochordamine A and B, which were isolated from ascidians as larval metamorphosis promoters, were converted to more polar compounds, urochordamine A and B, respectively, when left standing in protic solvents. These four compounds promoted larval metamorphosis of the ascidianHalocynthia roretzi in the order A>A>B>B, and induced metamorphosis of the pediveliger larvae of the musselMytilus edulis galloprovincialis.     

Sperm enzyme activity analysis in individual sperm for detection of heritable mutations in mammals

Summary Male mice were injected i.p. with 2.5 mg/kg mitomycin C, 100 mg/kg ethyl nitrosourea or saline and mated with untreated virgin females five weeks later. Sperm from 64 of the F₁ male progeny were analyzed histochemically for acrosin, succinic dehydrogenase and alpha-glycerophosphate dehydrogenase activity. The frequency of F₁ males with sub-normal sperm enzyme activity was significantly higher among progeny from treated males than in controls. These results show that analysis of sperm enzyme activity in F₁ males is a practical method for detection of transmitted mutations induced in a treated parent.We gratefully acknowledge USPHS, NIEHS grant 1 RO1 ES02607-02 and technical assistance by G. M. Oldford.     

Sperm predence in a hermaphroditic nematode (Caenorhabditis elegans) is due to competitive superiority of male sperm

When male and hermaphroditeCaenorhabditis elegans mate, the male's sperm outcompete the hermaphrodite's own sperm and fertilize a majority of the offspring. Here, we investigate the mechanism of male sperm precedence. We rule out the possibility that male sperm are stronger and more competitive because they are activated later than hermaphrodite sperm. We also find that a previously known gender difference in sperm activation does not influence sperm competition. Male sperm, rinsed free of seminal fluid, retained the capacity to take precedence after artificial insemination. Therefore, we conclude that male sperm themselves are competitively superior to hermaphrodite sperm. This trait maximizes outcrossing after mating and may increase both genetic diversity and heterozygosity of offspring whose parents, due to self-fertilization, may be highly homozygous.     

Localization of vitelline-coat lysin purified from testis of a top shell,Turbo cornutus

Summary The vitelline-coat lysin purified from the testis ofTurbo cornutus was found, by an immunofluorescence technique, to be located in the acrosome of the sperm, which suggested that the lysin reacts with the vitelline-coat in an early phase of fertilization to allow the sperm to penetrate through the coat.We are grateful to Professor M. Akino (Tokyo Metropolitan University) for his helpful advice.     

On the presence of different histidine decarboxylating enzymes in mammalian tissues

Zusammenfassung Säugetiergewebe enthalten mindestens zwei verschiedene, Histidin-decarboxylierende Fermente. Das eine, wahrscheinlich mit Dopadecarboxylase identisch, scheint für die Histaminbildung unter physiologischen Bedingungen ohne Bedeutung zu sein. Das andere ist in Mastzellen und wachsenden Geweben lokalisiert.

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This work has been supported by grants from the Swedish Medical Research Council.     

Manipulating the activity of immobilized enzymes with different organo-smectite complexes

Summary Enzymes may be immobilized on hydrophobic surfaces of organo-smectite complexes. The immobilized enzyme may be active, partially active, or inactive depending on the nature of the organic surface. These materials may be useful as models for enzymes in natural systems, and in medicine and industry.This work was partially supported by the National Science Foundation under Grant No. CHE-8306583. Journal Article No. 11603 of the Michigan Agricultural Experiment Station.     

Presence of 3,4-dihydroxyphenylalanine-containing peptides in hemocytes of the ascidian,Halocynthia roretzi

Summary Five 3,4-dihydroxyphenylalanine (DOPA)-containing peptides have been isolated from hemocytes of the ascidian,Halocynthia roretzi. Three of them were composed of DOPA, proline, phenylalanine, histidine and arginine in different ratios, while the other two contained only DOPA and an unidentified amino acid. DOPA-containing peptides were found to exist in only one type of hemocyte.     

Hormonal facilitation in the release of sperm from the spermatheca of the red-spotted newt

Summary Several neurotransmitters and hormones with potential to trigger a simultaneous contraction of the oviducts and the spermathecal myoepithelium were examined. Saline (0.05 ml), or 0.05 ml saline plus acetylcholine (9 mg), norepinephrine (50 g), arginine-vasotocin (25 units), prostaglandin F₂(3g) were injected into the spermathecal region of female newts (n=24 per group). The numbers of sperm present in the cloacae of prostaglandin-injected animals (107±30 SEM) were significantly greater than the numbers detected in saline (27±5 SEM) and in uninjected (14±3 SEM) controls. Smaller and less consistent increases in the numbers of sperm were detected in the vasotocin- and norepinephrine-injected groups. Study of sections from ovulating female newts failed to produce evidence that pressure from the passage of ova through the posterior portion of the oviduct forced sperm from the spermatheca. Observations indicate an active role for the spermathecal myoepithelium in the discharge of stored sperm and of a role for prostaglandin F₂ in triggering that discharge.     

Intracellular enzymes in renal lymph as a measure of anoxic injury of the kidney

Zusammenfassung Nachweis, dass 30-minutige Nierenischämie zum Austritt von Enzymen des Zellplasmas aus den Zellen führt. Nach 120 min Nierenischämie kommt es auch zum Austritt von lysosomalen und mitochondrialen Enzymen aus den Zellen.     

Activity of phospholipases A and some lysosomal enzymes in rat testes at different stages of hypervitaminosis A

Summary Excess of vitamin A induces decrease of neutral phospholipase A₁ and A₂ activity in rat testes homogenates on the 4th day, and increase of -galactosidase activity on the 8th day of treatment. It is suggested that phospholipase A activity decrease is of great importance in development of testicular disorders, caused by disbalance of vitamin A.     

Activity alterations of metabolic enzymes in the anterior pituitary of female rats during acute and chronic starvation,as well as after refeeding

Summary The activity of glycolysis and hexose monophosphate shunt decreases while the activity of some oxydative enzymes and acid phosphatase increases in the anterior pituitary of adult female rats during starvation. The alterations depend on the severity of starvation. The polypeptide hormone production also decreases. A close relationship exists between the metabolic activity of the gland and its endocrine function.     

Cyclic CMP (cytidine 3′,5′-monophosphate) suppresses changes in human sperm amplitude of lateral head displacement and hyperactivation

Summary The dibutyryl analog of cCMP suppressed sperm amplitude of lateral head displacement and hyperactivation. Sperm motility was inhibited by dibutyryl cCMP with a shift toward less linear trajectory sperm movements. The results suggest a role of cCMP as an inhibitory signal on sperm motility patterns related to sperm capacitation.     

Toxic effects of methylmercury on spermatozoa in vitro

Summary In an in vitro investigation, methylmercury (MeHg) reduced the motility of rat spermatozoa probably by the inhibition of succinate dehydrogenase and ATPase activities. Concomitant morphological changes observed in the spermatozoa were coiled tails and kinks in midpiece and tail regions.