Limits...
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.Prostasome-like vesicles have been isolated from pig seminal plasma by high-speed centrifugation and Sephadex G-200 gel chromatography.However, the two major bands were observed at approximately 90 kDa and approximately 60 kDa.

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

Fluorescence pattern of pig spermatozoa stained with FITC-PNA + PI for the assessment of acrosome status and sperm viability. Dead cells showing nuclear red PI fluorescence: C-D. Live cells without PI staining: A acrosome-reacted cells with uniform green FITC-PNA fluorescence of acrosome cap; B: acrosome-unreacted cells with no staining of acrosomal cap (original magnification ×1000).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2266753&req=5

Figure 4: Fluorescence pattern of pig spermatozoa stained with FITC-PNA + PI for the assessment of acrosome status and sperm viability. Dead cells showing nuclear red PI fluorescence: C-D. Live cells without PI staining: A acrosome-reacted cells with uniform green FITC-PNA fluorescence of acrosome cap; B: acrosome-unreacted cells with no staining of acrosomal cap (original magnification ×1000).

Mentions: Figure 4 shows a representative fluorescence pattern of pig spermatozoa stained with FITC-PNA + PI for the assessment of acrosome status and sperm viability


Prostasome-like vesicles stimulate acrosome reaction of pig spermatozoa.

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

Fluorescence pattern of pig spermatozoa stained with FITC-PNA + PI for the assessment of acrosome status and sperm viability. Dead cells showing nuclear red PI fluorescence: C-D. Live cells without PI staining: A acrosome-reacted cells with uniform green FITC-PNA fluorescence of acrosome cap; B: acrosome-unreacted cells with no staining of acrosomal cap (original magnification ×1000).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Fluorescence pattern of pig spermatozoa stained with FITC-PNA + PI for the assessment of acrosome status and sperm viability. Dead cells showing nuclear red PI fluorescence: C-D. Live cells without PI staining: A acrosome-reacted cells with uniform green FITC-PNA fluorescence of acrosome cap; B: acrosome-unreacted cells with no staining of acrosomal cap (original magnification ×1000).
Mentions: Figure 4 shows a representative fluorescence pattern of pig spermatozoa stained with FITC-PNA + PI for the assessment of acrosome status and sperm viability

Bottom Line: The presence of small membranous particles characterizes the male genital fluids of different mammalian species.Prostasome-like vesicles have been isolated from pig seminal plasma by high-speed centrifugation and Sephadex G-200 gel chromatography.However, the two major bands were observed at approximately 90 kDa and approximately 60 kDa.

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