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Query: UNIPROT:P56851 (epididymal)
 11,273 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Free Ca2+ changes the curvature of epididymal rat sperm flagella in demembranated sperm models. The radius of curvature of the flagellar midpiece region was measured and found to be a continuous function of the free Ca2+ concentration. Below 10(-7) M free Ca2+, the sperm flagella assumed a pronounced curvature in the same direction as the sperm head. The curvature reversed direction at 2.5 x 10(-6) M Ca2+ to assume a tight, hook-like bend at concentrations of 10(-5) to 10(-4) M free Ca2+. Sodium vanadate at 2 x 10(-6) M blocked flagellar motility, but did not inhibit the Ca2+-mediated change in curvature. Nickel ion at 0.2 mM and cadmium ion at 1 microM interfered with the transition and induced the low Ca2+ configuration of the flagellum. The forces that maintain the Ca2+-dependent curvature are locally produced, as dissection of the flagella into segments did not significantly alter the curvature of the excised portions. Irrespective of the induced pattern of curvature, the sperm exhibited coordinated, repetitive flagellar beating in the presence of ATP and cAMP. At 0.3 mM ATP the flagellar waves propagated along the principal piece while the level of free Ca2+ controlled the overall curvature. When Ca2+-treated sperm models with hooked midpieces were subjected to higher concentrations of ATP (1-5 mM), some cells exhibited a pattern of movement similar to hyperactivated motility in capacitated live sperm. This type of motility involved repetitive reversals of the Ca2+-induced bend in the midpiece, as well as waves propagated along the principal piece. The free Ca2+ available to the flagellum therefore appeared to modify both the pattern of motility and the flagellar curvature.
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PMID:Calcium regulation of flagellar curvature and swimming pattern in triton X-100--extracted rat sperm. 318 Feb 54

Binding sites for Con A and WGA were detected on bovine spermatozoa during epididymal maturation. We used colloidal gold as an EM-marker. The spermatozoa were treated according to a two-step method for lectin and colloidal gold, then adsorbed to lysine-coated nickel grids and subsequently examined by TEM in toto. Using this method we rapidly got information about the topographic distribution of lectin-binding sites. Major differences exist for WGA between caput and cauda spermatozoa. Conceding that cell-thickness poses some limitation, we consider this method to be practical and especially useful in studies concerning topographic distribution of cell surface components in single cell systems.
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PMID:Con A- and WGA-binding sites on bovine epididymal spermatozoa: TEM of specimens in toto. 623 81

Some characteristic features of the functional innervation of guinea-pig vas deferens have been determined. Both ganglionic transmission from the hypogastric nerves and impulse propagation in proximal regions (main branch bundles within about 15 mm from the prostatic end of the organ) of the majority of single postganglionic sympathetic fibres of vas deferens nerve, had a high safety factor. Failure at these levels cannot account for the intermittent pattern of electrically-evoked secretion of transmitter from the individual varicosity of the terminals of vas deferens nerves, observed under identical experimental conditions. The shape of the extracellular single fibre action potential recorded by small calibre suction electrodes remained constant in proximal regions of vas deferens nerve, when the frequency of stimulation was varied between 0.5 and 8 Hz. Therefore, frequency-dependent facilitation of transmitter secretion in this tissue cannot be explained by frequency-dependent growth in the amplitude of nerve action potentials, as earlier assumed. However, when recordings were made in distal regions of vas deferens nerve (in small axon bundles, close to their points of insertion into the substance of the epididymal end of the organ), on two occasions fibres were found in which the safety factor for impulse conduction was low and frequency-dependent. The possibility is discussed that this feature, which was an exception in these non-terminal regions of vas deferens nerve, may be shared by the majority of fibres as they proceed distally towards the terminals. Clearly, if this is the case, intermittent failure of transmitter secretion from the individual varicosity may be due, at least in part, to intermittent failure of conduction of the nerve impulse to the terminals. Some useful qualitative information on the ionic basis of the extracellular nerve action potential, that might underlie a proximo-distal decline in the safety factor for impulse conduction in these nerves, was obtained by determining the effects on the shape of the signal, caused by varying the ionic composition of the medium (sodium, calcium), and by local addition of agents with known actions on sodium (tetrodotoxin), potassium (tetra-ethyl ammonium, 4-aminopyridine, rubidium, barium) and calcium channels (cobalt, manganese, lanthanum, nickel, D-600). By these criteria, the action potential that was shown to be a "normal" sodium-potassium spike, in proximal regions of vas deferens nerve, was found to have a different "pharmacological profile", in distal regions of the nerve, in a manner suggesting that here nerve impulse conduction had become somehow "calcium-dependent".(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Frequency dependent intermittency and ionic basis of impulse conduction in postganglionic sympathetic fibres of guinea-pig vas deferens. 632 28

The 600-MHz proton spectrum of dethiobiotin (prepared from d-biotin with Raney nickel) was measured in order to gain information pertaining to its stereochemical homogeneity. The spectrum demonstrated clearly that the material is a 6:1 mixture of two stereoisomers. The cis compound, corresponding to the stereochemistry of d-biotin, is the major isomer. Two biotinyl- and two dethiobiotinylinsulins were prepared in which the distance between the biotins and insulin was varied by interposition of spacer arms. The synthesis of these compounds involved repeated N-hydroxysuccinimido ester condensations. Biotin N-hydroxysuccinimido ester, dethiobiotin N-hydroxysuccinimido ester, 6-aminohexanoic acid, and N-[3-[(3-aminopropyl)carboxyamino]-propyl]succinamic acid N-tert-butyl ester served as the building blocks for the spacers. The latter compound was prepared from N-[3-[(3-aminopropyl)amino]propyl]succinamic acid sulfate by the use of a selective amino-protecting method based on the differential stability toward acid of citraconyl and tert-butoxycarbonyl amino-protecting groups. The structure of N-[3-[(3-aminopropyl)amino]propyl]succinamic acid sulfate was established unequivocally by X-ray diffraction. The attachment of the biotinylated spacers to the insulin was exclusively at the N alpha, B1 position. Homogeneity of the final products as well as of the intermediates used in their synthesis was established by thin-layer chromatography, by high-pressure liquid chromatography, and in most instances by elemental analysis. The ratio of 6-aminohexanoic acid to lysine in hydrolysates of the insulin derivatives was in agreement with theory. The insulin derivatives were required for a study on the effect of avidin on their ability to interact with insulin receptors on rat epididymal adipocytes, which is described in the accompanying paper.
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PMID:Syntheses of biotinylated and dethiobiotinylated insulins. 638 May 70

Direct protein extraction from animals is the only approach available to obtain caltrin, calcium transport inhibitor. Here we report the expression and purification of caltrin, previously shown to hinder the influx of calcium into epididymal spermatozoa. Cloning of the caltrin gene into the pCDNA3.1 V5/His-TOPO vector and the subsequent ligation of the caltrin-His sequence into the transfer vector pBacPAK9 allowed the expression of recombinant caltrin using the baculovirus expression vector system (BEVS). Recombinant His-tagged caltrin was purified utilising both nickel (II)-nitrilotriacetic acid (Ni(2+)-NTA) and cobalt (II)-carboxymethylaspartate (Co(2+)-CmAsp) immobilised metal affinity chromatography (IMAC). Using the BEVS, caltrin-His was identified in the supernatant and in the cell lysate, suggesting that caltrin is a secreted protein. Based on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot results, purified recombinant caltrin-His was ascertained to be approximately 14.5kDa. Purification under the Co(2+) system yielded significantly purer protein samples when compared to the Ni(2+) system. Furthermore, Co(2+) was observed to bind the recombinant caltrin-His protein with higher efficiency and specificity and to yield a higher total protein concentration. Collectively, our results indicate that the Co(2+) system would be a better approach for purifying caltrin-His proteins than the Ni(2+).
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PMID:Expression of caltrin in the baculovirus system and its purification in high yield and purity by cobalt (II) affinity chromatography. 1276 21

Early studies on nickel essentiality with rats and goats indicated that nickel deprivation impaired reproductive performance. Nickel also has been found to influence cyclic nucleotide gated channels (CNG); these types of channels are important in sperm physiology. Thus, two experiments were conducted to test the hypothesis that nickel deficiency affects sperm physiology in a manner consistent with nickel having an essential function related to CNG channel functions. The experiments were factorially arranged with four treatment groups of eight weanling rats in each. In experiment 1, the treatments were supplemental dietary nickel of 0 and 1 mg/kg and N(omega)-nitro-L-arginine methyl ester (L-NAME, a nitric oxide synthase inhibitor) added to the drinking water (50 mg/100 mL) the last 3 wk of an 8-wk experiment. In experiment 2, the treatments were supplemental dietary nickel at 0 and 1 mg/kg and supplemental dietary sodium chloride (NaCl) at 0 and 80 g/kg. The NaCl and L-NAME variables were included to act as stressors affecting CNG channel activity. The basal diet contained per kilogram about 27 microg of nickel and 1 g of sodium. After 8 wk in experiment 1 and 16 wk in experiment 2, urine while fasting and testes and epididymis in both experiments, and seminal vesicles and prostates in experiment 2 were harvested for analysis. Nickel deprivation significantly decreased spermatozoa motility and density in the epididymides, epididymal transit time of spermatozoa, and testes sperm production rate. Nickel deficiency also significantly decreased the weights of the seminal vesicles and prostate glands. Excessive NaCl had no effect on sperm physiology; however, it decreased prostate gland weights. The findings support the hypothesis that nickel has an essential function that possibly could affect reproductive performance in higher animals, perhaps through affecting a CNG channel function.
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PMID:Nickel deficiency diminishes sperm quantity and movement in rats. 1283 98

Oxidative stress (OS) mechanisms are speculated to play a significant role in nickel-induced toxic effects and their carcinogenic potency. Although nickel-induced oxidative damage in somatic tissues is well demonstrated, evidence of the involvement of a similar mechanism(s) in nickel-induced testicular dysfunction and associated genotoxic effects is scarce. Hence, the present study aimed to investigate the nickel-induced OS response in testis and the associated genotoxic implications in vivo. Initially, the toxicity profile of nickel chloride was determined in adult albino mice (CFT-Swiss) following administration (intraperitoneal) of single doses. Subsequently, multiple sublethal doses (1.25, 2.5, and 5.0 micromol/100 g of body weight per day for 3 days) were used to characterize effects on testicular histoarchitecture, lipid peroxidation (LPO) in testis (homogenates, microsomal or mitochondrial fractions) and epididymal sperm, DNA damage, induction of apoptosis in testis, and incidence of sperm head abnormalities. Although short-term doses of nickel induced only a minimal LPO response, multiple doses elicited a moderate (15% to 30%) increase in LPO in whole homogenates and higher dose-related increases in both mitochondrial (20% to 50%) and microsomal fractions (25% to 60%). This was associated with a significant increase in DNA damage in the testis as evidenced by increased single-strand breaks (fluorimetric analysis of DNA unwinding assay). Further, at higher doses, nickel-induced apoptosis was demonstrable in the testis biochemically. Although caudal sperm counts determined at all sampling weeks showed no alterations, analysis for head abnormalities revealed a nearly 3- to 4-fold increase in the percentage of abnormal sperms among the nickel-treated males during the first 3 weeks. Furthermore, mating of nickel-treated (2.5 micromol/100 g of body weight per day for 5 days) males sequentially for a period of 5 weeks with untreated females resulted in a significant increase in male-mediated dominant lethal-type mutations (the frequency of dead implantations) during the first 3 weeks, suggesting a stage-specific effect on postmeiotic germ cells. These findings suggest that testicular toxicity of nickel compounds may be related to enhanced production of reactive oxygen species, probably mediated through oxidative damage to macromolecules, including damage to DNA.
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PMID:Nickel-induced oxidative stress in testis of mice: evidence of DNA damage and genotoxic effects. 1547 75

Improper management of hazardous wastes adversely impacts the environment and the public health. The present study was aimed at investigating the influence of Olushosun municipal landfill leachate (OMLL) from Ojota in the Lagos State of Nigeria on testicular function by assessing the plasma concentrations of reproductive hormones, testicular biometal levels, and antioxidant levels as well as observing the histological alterations in testes and epididymides of rats after exposure to 0, 12.5, and 25% OMLL in drinking water for 7 days. Exposure to OMLL significantly decreased the daily fluid intake, but it resulted in testicular biometal accumulation as follows: lead > cadmium > nickel > iron > copper. Acute exposure to OMLL induced oxidative stress and increased the activities of marker enzymes of testicular function but markedly decreased the circulatory concentrations of luteinizing hormone, follicle-stimulating hormone, prolactin, testosterone, thyroid-stimulating hormone, triiodothyronine, and thyroxine. Testicular and epididymal degeneration with significant decrease in sperm quality and quantity were observed in OMLL-exposed rats. Collectively, the data presented herein indicate that exposure to OMLL-induced testicular dysfunction associated with biometal accumulation and endocrine disruption in rats. If the effects can be extrapolated to humans, OMLL may present significant health implications for individuals exposed to OMLL-contaminated substances.
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PMID:Municipal landfill leachate-induced testicular oxidative damage is associated with biometal accumulation and endocrine disruption in rats. 2517 71