Effect of Cryopreservation on Sea Bass Sperm Proteins

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...

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Bibliographic Details
Published in:Biology of reproduction 2005-05, Vol.72 (5), p.1262-1267
Main Authors: Zilli, Loredana, Schiavone, Roberta, Zonno, Vincenzo, Rossano, Rocco, Storelli, Carlo, Vilella, Sebastiano
Format: Article
Language:English
Subjects:
Animals
Bass - metabolism
Biological and medical sciences
Contents
Cryopreservation
Danio rerio
Dicentrarchus labrax
Electrophoresis, Gel, Two-Dimensional
fertilization
Fish Proteins - isolation & purification
Fish Proteins - metabolism
Fundamental and applied biological sciences. Psychology
gamete biology
Male
Mammalian male genital system
Morphology. Physiology
Peptide Mapping
Semen Preservation
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
sperm
Spermatozoa - metabolism
Vertebrates: reproduction
Xenopus Proteins - isolation & purification
Zebrafish Proteins - isolation & purification
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Summary: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.
ISSN:0006-3363
1529-7268
DOI:10.1095/biolreprod.104.036202