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Prostasome-like vesicles stimulate acrosome reaction of pig spermatozoa.

Siciliano L, Marcianò V, Carpino A - Reprod. Biol. Endocrinol. (2008)

Bottom Line: The presence of small membranous particles characterizes the male genital fluids of different mammalian species.However, the two major bands were observed at approximately 90 kDa and approximately 60 kDa.A vesicle-mediated transfer of aminopeptidase to sperm cells has been also detected.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell Biology, Faculty of Pharmacy, University of Calabria, Cosenza, Italy. siciliano_laura@yahoo.it

ABSTRACT

Background: The presence of small membranous particles characterizes the male genital fluids of different mammalian species. The influence of semen vesicles, denominated prostasomes, on sperm functional properties has been well documented in humans, but their biological activity is scarcely known in other species. The present work investigated prostasome-like vesicles in pig semen for their ability to interact with spermatozoa and to affect acrosome reaction.

Methods: Prostasome-like vesicles have been isolated from pig seminal plasma by high-speed centrifugation and Sephadex G-200 gel chromatography. Morphology of purified vesicles has been checked by scanning electron microscopy while their protein pattern has been investigated by SDS-PAGE. Then prostasome- like vesicles have been incubated with pig spermatozoa and their ability to interact with sperm has been tested by the aminopeptidase assay. In addition, the efficiency of vesicles to influence the acrosome reaction has been investigated by assessing the sperm acrosomal status by the PI/FITC-PNA (propidium iodide/fluorescein isothiocyanate-labeled peanut agglutinin) stainings.

Results: Purified vesicles revealed a complex protein pattern with the occurrence of bands in the high, medium and low molecular weight range. However, the two major bands were observed at approximately 90 kDa and approximately 60 kDa. A vesicle-mediated transfer of aminopeptidase to sperm cells has been also detected. Furthermore, a significant increase of acrosome reaction extent has been revealed in spermatozoa incubated with prostasome-like vesicles in comparison to control sperm.

Conclusion: This is the first report demonstrating that pig prostasome-like vesicles are able, in vitro, to interact with spermatozoa and to stimulate the acrosome reaction. These findings lead to hypothesize a transfer of molecules from vesicles to sperm membrane, thus sensitizing male gametes to undergo the acrosome reaction.

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SDS-PAGE: Protein separation from 30 μg of pig purified prostasome-like vesicles on a Coomassie stained gel. Lane 1: molecular weight standards Lane 2: prostasome like-vesicles.
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Figure 2: SDS-PAGE: Protein separation from 30 μg of pig purified prostasome-like vesicles on a Coomassie stained gel. Lane 1: molecular weight standards Lane 2: prostasome like-vesicles.

Mentions: Following the SDS-PAGE protein separation, all the samples have revealed a complex protein pattern. Six distinct bands were observed in the high molecular weight range, with two major bands approximatively at 90 kDa and 60 kDa. In addition, some consistent bands were also detected in the medium and low molecular weight range (approximatively at 41, 31, 28, 25, 23, 19, 16 and 6 kDa). Figure 2 shows a Coomassie-stained SDS-PAGE gel of separated proteins from one representative sample


Prostasome-like vesicles stimulate acrosome reaction of pig spermatozoa.

Siciliano L, Marcianò V, Carpino A - Reprod. Biol. Endocrinol. (2008)

SDS-PAGE: Protein separation from 30 μg of pig purified prostasome-like vesicles on a Coomassie stained gel. Lane 1: molecular weight standards Lane 2: prostasome like-vesicles.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2266753&req=5

Figure 2: SDS-PAGE: Protein separation from 30 μg of pig purified prostasome-like vesicles on a Coomassie stained gel. Lane 1: molecular weight standards Lane 2: prostasome like-vesicles.
Mentions: Following the SDS-PAGE protein separation, all the samples have revealed a complex protein pattern. Six distinct bands were observed in the high molecular weight range, with two major bands approximatively at 90 kDa and 60 kDa. In addition, some consistent bands were also detected in the medium and low molecular weight range (approximatively at 41, 31, 28, 25, 23, 19, 16 and 6 kDa). Figure 2 shows a Coomassie-stained SDS-PAGE gel of separated proteins from one representative sample

Bottom Line: The presence of small membranous particles characterizes the male genital fluids of different mammalian species.However, the two major bands were observed at approximately 90 kDa and approximately 60 kDa.A vesicle-mediated transfer of aminopeptidase to sperm cells has been also detected.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cell Biology, Faculty of Pharmacy, University of Calabria, Cosenza, Italy. siciliano_laura@yahoo.it

ABSTRACT

Background: The presence of small membranous particles characterizes the male genital fluids of different mammalian species. The influence of semen vesicles, denominated prostasomes, on sperm functional properties has been well documented in humans, but their biological activity is scarcely known in other species. The present work investigated prostasome-like vesicles in pig semen for their ability to interact with spermatozoa and to affect acrosome reaction.

Methods: Prostasome-like vesicles have been isolated from pig seminal plasma by high-speed centrifugation and Sephadex G-200 gel chromatography. Morphology of purified vesicles has been checked by scanning electron microscopy while their protein pattern has been investigated by SDS-PAGE. Then prostasome- like vesicles have been incubated with pig spermatozoa and their ability to interact with sperm has been tested by the aminopeptidase assay. In addition, the efficiency of vesicles to influence the acrosome reaction has been investigated by assessing the sperm acrosomal status by the PI/FITC-PNA (propidium iodide/fluorescein isothiocyanate-labeled peanut agglutinin) stainings.

Results: Purified vesicles revealed a complex protein pattern with the occurrence of bands in the high, medium and low molecular weight range. However, the two major bands were observed at approximately 90 kDa and approximately 60 kDa. A vesicle-mediated transfer of aminopeptidase to sperm cells has been also detected. Furthermore, a significant increase of acrosome reaction extent has been revealed in spermatozoa incubated with prostasome-like vesicles in comparison to control sperm.

Conclusion: This is the first report demonstrating that pig prostasome-like vesicles are able, in vitro, to interact with spermatozoa and to stimulate the acrosome reaction. These findings lead to hypothesize a transfer of molecules from vesicles to sperm membrane, thus sensitizing male gametes to undergo the acrosome reaction.

Show MeSH
Related in: MedlinePlus