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BOR - Papers in Press, published online ahead of print January 19, 2005.
Biol Reprod 2005, 10.1095/biolreprod.104.036202
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BIOLOGY OF REPRODUCTION 72, 1262–1267 (2005)
DOI: 10.1095/biolreprod.104.036202
© 2005 by the Society for the Study of Reproduction, Inc.

Effect of Cryopreservation on Sea Bass Sperm Proteins

Loredana Zilli 2, Roberta Schiavone 2, Vincenzo Zonno 3, Rocco Rossano 4, Carlo Storelli 2, and Sebastiano Vilella 1 2

Laboratorio di fisiologia comparata, Dipartimento di Scienze e Tecnologie biologiche ed ambientali,2University of Lecce, 73100 Lecce, Italy Marine Aquaculture and Fisheries Research Centre (M.A.R.C.),3 73100 Lecce, Italy Dipartimento di Biologia,4 Difesa e Biotecnologie Agro-Forestali, University of Basilicata, 85100 Potenza, Italy

In the present study we used two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to verify whether the protein expression of sea bass sperm was affected by the cryopreservation procedure. The protein profiles differed between fresh and frozen-thawed semen as revealed by visual inspection and by image analysis software. We identified 163 spots in fresh sperm; among these, 13 were significantly decreased and 8 were absent in two-dimensional gel obtained with cryopreserved sperm. Five of these spots were analyzed with MALDI-TOF, but only three showed a significant match in the databases used in bio-informatics analysis (PeptIdent, Mascot, and MS-Fit). In particular, spot 5 showed homology with a novel protein of zebrafish (similar to SKB1 of human and mouse), spot 13 showed homology with amphibian G1/S-specific cyclin E2, and spot 20 showed homology with the hypothetical protein DKFZp566A1524 of Brachidanio rerio. The present work shows that the use of the cryopreservation procedure causes the degradation of sperm proteins and among these, two could be at least partially responsible for the observed decrease in sperm motility duration and the lower hatching rate of eggs fertilized with cryopreserved sperm.

gamete biology, fertilization, sperm

1 Correspondence: Vilella Sebastiano, Laboratorio di Fisiologia Comparata, Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali University of Lecce, Via Provinciale Lecce-Monteroni, 73100 Lecce, Italy. FAX: 39 083 232 4220; sebastiano.vilella{at}unile.it






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Copyright © 2005 by the Society for the Study of Reproduction.