Purification and characterization of β‐glucuronidase from bull seminal plasma and its role in fertilization

Rethinaswamy, Asokan; Yang, Chung-May; Srivastava, Pranay

doi:10.1002/mrd.1080380408

Cited by 6 publications

(2 citation statements)

References 32 publications

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“…1996). This enzyme has been shown to act on glycosaminoglycans, and together with hyaluronidase, it is involved in dispersing cumulus and corona cells (Rethinaswamy et al. 1994).…”

Section: Discussionmentioning

confidence: 99%

Alkaline and Acid Phosphatase, β‐Glucuronidase and Electrolyte Levels in Fractionated Stallion Ejaculates

Kareskoski

Reilas

Sankari

et al. 2010

Reprod Domestic Animals

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Seminal plasma (SP) is a mixture of contents from the testes, epididymides and accessory sex glands. The sperm concentration is highest in the first few jets, or fractions, of the ejaculate, and the composition of SP varies between these fractions because accessory gland secretions are released in a specific order. The aim of this study was to compare the levels of Na, Cl, K, Mg, Ca, inorganic phosphate (Pi) and the enzymes alkaline phosphatase (AP), acid phosphatase (ACP) and β-glucuronidase (BG) in the different fractions of the ejaculate and in different stallions. All semen collections were done using a computer-controlled phantom that collects the ejaculatory jets separately in five cups. The cups with the highest (HIGH) and the lowest (LOW) sperm concentration were analysed. In Trial I, semen was collected from three reproductively normal stallions. In Trial II, ejaculates of two reproductively normal stallions were compared to those of two subfertile stallions. In Trial III, semen was collected from seven stallions with varying reproductive history. The sperm-rich fractions contained the highest levels of AP, ACP, BG and inorganic phosphate, and the values were positively correlated to the sperm concentration. Significant differences between the subfertile and the fertile stallions pairs in HIGH:LOW ratios were found in Pi and Cl concentrations. The highest concentrations of Ca and Mg were found in the last fractions with low sperm concentrations, with no significant differences between the fertile and the subfertile stallion pairs. The concentrations of K, Na and Cl were similar in HIGH and LOW fractions and in whole ejaculate samples. Pre-sperm fluid contained the highest concentrations of Na and Cl. Some of the possible variation in storage tolerance between ejaculates and ejaculatory fractions could perhaps be explained by differences in the composition of SP.

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“…1996). This enzyme has been shown to act on glycosaminoglycans, and together with hyaluronidase, it is involved in dispersing cumulus and corona cells (Rethinaswamy et al. 1994).…”

Section: Discussionmentioning

confidence: 99%

Alkaline and Acid Phosphatase, β‐Glucuronidase and Electrolyte Levels in Fractionated Stallion Ejaculates

Kareskoski

Reilas

Sankari

et al. 2010

Reprod Domestic Animals

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“…Dias et al (2004) identified α-1,4-glucosidase activity in stallion seminal plasma and epididymis with increasing activity from the proximal caput to the cauda. Among these seminal plasma glycosidases, at least BGase and Nacetyl-β-D-glucosaminidase (NAGase) bind with high affinity to the sperm surface and may play a part in modifying the sperm surface before fertilization (Barbieri et al 1996) and cause dispersion of the cumulus oophorus cells (Rethinaswamy et al 1994). Levels of α-galactosidase, β-galactosidase and NAGase have been shown to differ significantly between infertile men with oligo-astheno-teratozoospermia and fertile controls (Corrales et al 2000).…”

Section: Seminal Plasma Enzymesmentioning

confidence: 99%

Components of stallion seminal plasma and their influence on spermatozoa

Katila¹,

Kareskoski²

2006

PHK

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SummaryThis review focuses on the composition of stallion seminal plasma and on its role as a storage medium of spermatozoa, with special reference to artificial insemination (AI). The stallion ejaculates in 6-9 jets resulting from urethral contractions, and these jets have characteristic volumes, sperm concentrations and composition. Different types of open-ended or closed artificial vaginas (AVs) are available for semen collection, and ejaculatory jets can be collected separately as a fractionated ejaculate, using an open-ended AV or an automated phantom (Equidame ® ). Seminal plasma influences spermatozoa during storage in different ways depending on dilution ratios, duration of storage and other factors. Beneficial effects have been reported after seminal plasma addition before freezing, but contradictory results show that short-term exposure (15 min) of sperm to seminal plasma has no effect on sperm motility or viability, and prolonged exposure (2, 4 or 6 h) to 5 or 20% seminal plasma in the freezing diluent is deleterious. The optimal proportion of seminal plasma for cooled storage seems to be fairly low. Figures from 0 to 20% have been reported in different studies. There is some evidence that the sperm-rich fractions of the ejaculate would be more suitable for freezing than the whole ejaculate or the sperm-low fractions. Seasonal and individual variation is seen in the composition and quality of stallion seminal plasma. Research on the biochemical composition of whole and fractionated stallion ejaculates, mainly concerning the levels of enzymes, carbohydrates, lipids, electrolytes and minerals, and compounds involved in the protection of sperm from oxidative damage is reviewed.Keywords: horse, reproduction, seminal plasma, spermatozoa, cryopreservation, cooled semen Zusammensetzung des Seminalplasmas beim Hengst und dessen Einfluss auf die SpermienDieser Übersichtsartikel befasst sich mit der Zusammensetzung des Seminalplasmas beim Hengst und dessen Bedeutung als Komponente in konserviertem Sperma unter besonderer Berücksichtigung der praktischen Anforderungen in der equinen Samenübertragung. Mithilfe urethraler Kontraktionen erfolgt die Ejakulation beim Hengst in 6-9 Fraktionen; diese Fraktionen unterscheiden sich hinsichtlich des Volumens, der Spermienkonzentration und der Zusammensetzung. Verschiedene Typen (geschlossene und offene Modelle) künstlicher Vaginen werden zur Samengewinnung eingesetzt. Die verschiedenen Ejakulatfraktionen können manuell -mit offenem Scheidenmodell -oder mithilfe eines automatisierten Phantoms (Equidame ® ) separat gewonnen werden. Seminalplasma beeinflusst die Spermien während der Lagerung in Abhängigkeit vom Verdünnungsgrad, der Lagerungsdauer und weiteren Faktoren. Positive Effekte auf die Spermaqualität wurden durch Seminalplasmazugabe vor der Tiefgefrierung des Samens erzielt; im Gegensatz dazu wird jedoch beschrieben, dass eine kurzzeitige Exposition (15 Min.) von Spermien mit Seminalplasma keine Effekte auf Spermienmotilität und -viabilität bedingt, eine prolongierte...

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?-D-Glucuronidase is associated with goat sperm cytoskeleton

et al. 2001

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Glycosidase activities were detected as detergent‐insoluble after sequential extractions of goat sperm with Triton X‐100. Seventy percent of total β‐glucuronidase activity was found in the detergent‐insoluble fraction. This portion of β‐glucuronidase was resistant to extractions in the presence of 1 M KCl, chaotropic agents, colchicine, or cytochalasin B, being only partially solubilized by 3 M KCl or DNAse I treatment. The treatment with 0.1% sodium deoxycholate was effective, releasing 73% of the enzyme activity. Treating the deoxycholate extract with DNAse I resulted in a change in the elution profile of β‐glucuronidase as judged by gel filtration chromatography. A polyclonal antibody was developed against pancreatic β‐glucuronidase, and the sperm enzyme was strongly inhibited by the IgG fraction of this antibody. Western blot analysis showed that the same protein correspond to both Triton‐soluble and insoluble enzyme. Results demonstrate that β‐glucuronidase is tightly bound to the Triton X‐100 resistant fraction, suggesting that the enzyme is associated to sperm cytoskeleton. J. Exp. Zool. 289:146–152, 2001. © 2001 Wiley‐Liss, Inc.

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Purification and characterization of β‐glucuronidase from bull seminal plasma and its role in fertilization

Cited by 6 publications

References 32 publications

Alkaline and Acid Phosphatase, β‐Glucuronidase and Electrolyte Levels in Fractionated Stallion Ejaculates

Alkaline and Acid Phosphatase, β‐Glucuronidase and Electrolyte Levels in Fractionated Stallion Ejaculates

Components of stallion seminal plasma and their influence on spermatozoa

?-D-Glucuronidase is associated with goat sperm cytoskeleton

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