Effects of putative protease inhibitors on the acrosome reaction of sea urchin spermatozoa

Hyne, Ross V.; Garbers, David L.

doi:10.1530/jrf.0.0660051

Cited by 5 publications

(3 citation statements)

References 24 publications

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“…The purpose of this study is the development of conditions that will facilitate the preservation of sperm attributes during overnight shipment of home‐collected semen samples to a central laboratory by men under investigation for reproductive toxicity exposure or infertility. The preserving agent, PMSF, is an enzyme inhibitor that has been used previously in animal semen (Hyne and Garbers, 1982; Frankel and Chapman, 1984; Imschenetzky et al, 1997). For the overnight incubations, we used conventional Falcon tubes or the Transem100 semen shipper (Fertility Solutions, Cleveland, Ohio) with no discernible differences.…”

Section: Resultsmentioning

confidence: 99%

“…The utility of PMSF, a proteolytic enzyme inhibitor, has been explored previously for the preservation of sperm nuclear proteins in sea urchins and rodents. Addition of PMSF preserved the fertilizing ability of sperm nuclei, and inhibited the degradation of sperm histones during pronuclear remodeling (Hyne and Garbers, 1982; Imschenetzky et al, 1997). The documented stabilization of sperm nuclear proteins might be relevant to the PMSF preservation of DNA integrity and the aniline blue chromatin staining pattern of human sperm reported in this study.…”

Section: Discussionmentioning

confidence: 99%

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Semen Characteristics After Overnight Shipping: Preservation of Sperm Concentrations, HspA2 Ratios, CK Activity, Cytoplasmic Retention, Chromatin Maturity, DNA Integrity, and Sperm Shape

Huszár

Çelik-Özenci

Çaylı

et al. 2004

Journal of Andrology

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ABSTRACT:We tested several approaches that can be used to preserve sperm attributes and the objective biochemical markers of sperm maturity and function for assessment in a remote centralized laboratory after overnight shipping of semen samples. Addition of phenyl-methyl-sulfonyl-fluoride (PMSF) to a final concentration of 20 g/mL semen at 4ЊC has preserved sperm concentrations and HspA2 isoform ratios, even at room temperature, simulating a shipping delay in moderate ambient temperatures. Regarding the attributes of individual spermatozoa, the patterns of CK-immunocytochemistry (demonstrates cytoplasmic retention in diminished-maturity spermatozoa); aniline blue staining pattern (tests chromatin maturity); sperm shape assessed by both Kruger strict morphology and computer assisted morphometry; and sperm DNA integrity, as tested by DNA nick translation, all remained unchanged. Thus, the PMSF-4ЊC conditions preserved sperm concentrations and the cytoplasmic and nuclear biomarkers of sperm cellular maturity and function for next-day analysis. This shipping method will facilitate the early detection of subtle changes in semen quality that can affect sperm function, even when there has been no decline in sperm concentrations to signal possible toxic effects. Furthermore, sample preservation will enable investigators to evaluate semen for toxicology studies and for diagnosis of male infertility from remote locations. Home collection of semen should enhance study participation, and semen assessment in centralized laboratories will address concerns regarding interlaboratory variations and quality control.Key words: Reproductive toxicity, male infertility, cytoplasmic and nuclear biomarkers.J Androl 2004;25:593-604 E xposures to environmental toxicants can have detrimental effects on human male reproduction (Schrader et al, 1992;Wyrobek et al, 1997). In field studies, two levels of semen analysis are usually carried out: primary studies, which can include the conventional semen parameters of sperm concentration, motility, and morphology, as well as measurement of specific biomarkers selected by the expected health effects of the environmental toxicants causing the exposure, and secondary studies based on archived samples (ie, frozen samples, videotaped microscopic fields, or sperm smears) that are used later to further evaluate specific findings. We are developing a potential third approach, which recognizes that each semen sample is composed of sperm populations of various levels of cellular maturity. The proportions of mature and diminished-maturity sperm, which define the functional efficacy of a semen sample, can be measured by objective biochemical methods, independently from sperm concentrations and motility. This approach provides two potential advantages: 1) reliable sperm biochemical measurements, which could reflect changes in fertility and in the paternal contribution of sperm DNA to zygote development, even when a man has not reached a level of exposure that would affect his sperm concentration, and 2) detec...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Semen Characteristics After Overnight Shipping: Preservation of Sperm Concentrations, HspA2 Ratios, CK Activity, Cytoplasmic Retention, Chromatin Maturity, DNA Integrity, and Sperm Shape

Huszár

Çelik-Özenci

Çaylı

et al. 2004

Journal of Andrology

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“…The acrosome reaction of sea-urchin spermatozoa requires extracellular Na+ (Schackmann, Eddy & Shapiro, 1978). It is inhibited by an increase in extracellular + (Schackmann et al, 1978), is pH dependent (Collins & Epel, 1977;Hyne & Garbers, 1982) and is also accompanied by uptake of 45Ca2+ and 22Na+, as well as release of K+ and H+ . Induction of the sea-urchin sperm acrosome reaction leads to depolarization of the membrane potential (Schackmann, Christen & Shapiro, 1981).…”

Section: Alteredmentioning

confidence: 99%

Sodium requirement for capacitation and membrane fusion during the guinea-pig sperm acrosome reaction

Hyne¹,

Higginson²,

Kohlman³

et al. 1984

Reproduction

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Guinea-pig spermatozoa, incubated in a minimal culture medium (MCM-PL) containing 150 mM-Na+ and no added K+, capacitate within 2 h and respond to added Ca2+ with maximal percentage of acrosome reactions. When spermatozoa, initially capacitated in Medium MCM-PL, were washed and resuspended in saline-based media, the motile spermatozoa showed acrosome reactions only in response to added Ca2+ at an extracellular pH greater than 7.4. In contrast, resuspension of capacitated spermatozoa into isotonic sucrose or choline-based media containing no added Na+ and K+ (pH 7.8) did not support acrosome reactions in response to the addition of Ca2+. Examination under the electron microscope showed that these spermatozoa responded to added Ca2+ with swelling or cavitation of the acrosomal matrix but fusion of the plasma membrane and acrosomal membrane did not occur. Treatment of spermatozoa with monensin, a monovalent cationic ionophore, induced rapid acrosome reactions in the presence of extracellular Ca2+ and Na+. In contrast, spermatozoa incubated for 3 h in a sucrose-based Na+-deficient medium or Medium MCM-PL containing 10 mM-KCl failed to give a significant percentage of acrosome reactions in response to the addition of Ca2+, but the addition of ouabain (0.1 mM) prevented the K+ inhibition. Amiloride, a sodium channel blocker in various other tissues, retarded the development of acrosome reactions of spermatozoa incubated in Medium MCM-PL. Based on these data, it appears that an increase in intracellular Na+ has a functional role in acquisition of guinea-pig sperm fertilizing ability.

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Expression of Na+/H+ exchanger isoforms 1, 2, 3, and 4 in bovine endometrium and the influence of uterine pH at time of fixed-time AI of pregnancy success

Bolzenius

Cushman

Perry

2016

Animal Reproduction Science

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Effects of putative protease inhibitors on the acrosome reaction of sea urchin spermatozoa

Cited by 5 publications

References 24 publications

Semen Characteristics After Overnight Shipping: Preservation of Sperm Concentrations, HspA2 Ratios, CK Activity, Cytoplasmic Retention, Chromatin Maturity, DNA Integrity, and Sperm Shape

Semen Characteristics After Overnight Shipping: Preservation of Sperm Concentrations, HspA2 Ratios, CK Activity, Cytoplasmic Retention, Chromatin Maturity, DNA Integrity, and Sperm Shape

Sodium requirement for capacitation and membrane fusion during the guinea-pig sperm acrosome reaction

Expression of Na+/H+ exchanger isoforms 1, 2, 3, and 4 in bovine endometrium and the influence of uterine pH at time of fixed-time AI of pregnancy success

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