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

Reproductive Biology and Endocrinology

Research

Gabriella Zito¹  Elisa Giacomini¹  Elena Giolo²  Stefania Luppi²  Monica Martinelli²  Giuseppe Ricci³  Rodolfo C Garcia⁴ 

[1] Department of Medicine, Surgery and Health Sciences, University of Trieste, 34149, Trieste, Italy;Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell’Istria 65/1, 34137, Trieste, Italy;Institute for Maternal and Child Health, IRCCS Burlo Garofolo, Via dell’Istria 65/1, 34137, Trieste, Italy;Department of Medicine, Surgery and Health Sciences, University of Trieste, 34149, Trieste, Italy;International Centre for Genetic Engineering and Biotechnology (I.C.G.E.B.), Area Science Park, 34149, Trieste, Italy;

关键词: Density gradient centrifugation;    Sperm cells;    Swim-up;    Protein 2-dimensional maps;   

DOI  :  10.1186/s12958-015-0027-y

 received in 2015-01-16, accepted in 2015-04-09,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundReports 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.MethodsProtein 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).ResultsNo 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.ConclusionsThe 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.

【 授权许可】

Unknown   
© Luppi et al.; licensee BioMed Central. 2015. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.