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The effect of reactive oxygen species on motility parameters, DNA integrity, tyrosine phosphorylation and phosphatase activity of common carp ( Cyprinus carpio L.) spermatozoa
Author(s) -
Gazo Ievgeniia,
ShaliutinaKolešová Anna,
Dietrich Mariola A.,
Linhartová Pavla,
Shaliutina Olena,
Cosson Jacky
Publication year - 2015
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.22442
Subject(s) - biology , sperm motility , tyrosine phosphorylation , xanthine oxidase , oxidative stress , protein tyrosine phosphatase , carp , reactive oxygen species , biochemistry , phosphatase , phosphorylation , sperm , motility , acid phosphatase , common carp , dna fragmentation , oxidative phosphorylation , microbiology and biotechnology , cyprinus , enzyme , programmed cell death , apoptosis , botany , fishery , fish <actinopterygii>
SUMMARY The effect of reactive oxygen species production on the motility parameters, DNA integrity, acid phosphatase activity, and protein tyrosine phosphorylation in spermatozoa of the common carp ( Cyprinus carpio ) was investigated. Spermatozoa were exposed to different concentrations of xanthine and xanthine oxidase (X‐XO) either in the presence or absence of antioxidants for 15 and 60 min. A dose‐ and time‐dependent reduction in spermatozoa motility and velocity was observed. Comet assays showed a dramatic increase in DNA fragmentation after 15 min. Changes in tyrosine phosphorylation of spermatozoa proteins were observed by Western blotting with anti‐phosphotyrosine antibodies, and proteins of interest were identified by mass spectrometry. After a 60 min exposure to X‐XO, O‐linked N‐acetylglucosamine transferase, isoform 4 was phosphorylated and septin‐8‐A was dephosphorylated. Acid phosphatase activity also decreased in a dose‐dependent manner after a 60 min exposure to oxidative stress. The results demonstrate that oxidative stress impaired functional variables (sperm motility, velocity, DNA integrity) of carp spermatozoa, and altered intracellular signalling pathways through changes in tyrosine phosphorylation and acid phosphatase activity. Mol. Reprod. Dev. 82: 48–57, 2015. © 2014 Wiley Periodicals, Inc .