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Reproductive Biology and Endocrinology
Published in: Reproductive Biology and Endocrinology 1/2015

Open Access 01-12-2015 | Research

Comparative proteomic analysis of spermatozoa isolated by swim-up or density gradient centrifugation

Authors: Stefania Luppi, Monica Martinelli, Elisa Giacomini, Elena Giolo, Gabriella Zito, Rodolfo C Garcia, Giuseppe Ricci

Published in: Reproductive Biology and Endocrinology | Issue 1/2015

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Abstract

Background

Reports about the morphologic and functional characteristics of spermatozoa prepared by density gradient centrifugation (DC) or swim-up (SU) have produced discordant results. We have performed a proteomic comparison of cells prepared by DC and SU providing a molecular insight into the differences between these two methods of sperm cell isolation.

Methods

Protein maps were obtained by 2-dimensional (2-D) separations consisting of isoelectrofocusing (IEF) from pI 3 to 11 followed by SDS-polyacrylamide gel electrophoresis. 2-D gels were stained with Sypro Ruby. Map images of DC and SU spermatozoa were compared using dedicated software. Intensities of a given spot were considered different between DC and SU when their group mean differed by >1.5-fold (p < 0.05, Anova).

Results

No differences were observed for 853 spots, indicating a 98.7% similarity between DC and SU. Five spots were DC > SU and 1 was SU > DC. Proteins present in 3 of the differential spots could be identified. One DC > SU spot contained lactate dehydrogenase C and gamma-glutamylhydrolase, a second DC > SU spot contained fumarate hydratase and glyceraldehyde-3-phosphate dehydrogenase-2, and a SU > DC spot contained pyruvate kinase M1/M2.

Conclusions

The differences in protein levels found on comparison of DC with SU spermatozoa indicate possible dissimilarities in their glycolytic metabolism and DNA methylation and suggest that DC cells may have a better capacitation potential.
Appendix
Available only for authorised users
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Metadata
Title
Comparative proteomic analysis of spermatozoa isolated by swim-up or density gradient centrifugation
Authors
Stefania Luppi
Monica Martinelli
Elisa Giacomini
Elena Giolo
Gabriella Zito
Rodolfo C Garcia
Giuseppe Ricci
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Reproductive Biology and Endocrinology / Issue 1/2015
Electronic ISSN: 1477-7827
DOI
https://doi.org/10.1186/s12958-015-0027-y

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