Association of the cysteine-rich secretory protein-3 (CRISP-3) and some of its polymorphisms with the quality of cryopreserved stallion semen
Alexandra Usuga A D , Benjamín A. Rojano B and Giovanni Restrepo CA Faculty of Veterinary Medicine and Animal Science, CES University, Cl. 10 No. 22-04, 050021, Medellín, Colombia.
B Faculty of Sciences, School of Chemistry, Universidad Nacional de Colombia, Cl. 59 A No. 63-20, 050034, Medellín, Colombia.
C Faculty of Agricultural Sciences, Department of Animal Science, Universidad Nacional de Colombia, Cl. 59 A No. 63-20, 050034, Medellín, Colombia.
D Corresponding author. Email: ausuga@ces.edu.co
Reproduction, Fertility and Development 30(3) 563-569 https://doi.org/10.1071/RD17044
Submitted: 7 February 2017 Accepted: 8 August 2017 Published: 31 August 2017
Abstract
Contribution of seminal plasma proteins to semen freezability has been reported in several species, suggesting these proteins as genetic markers. The aim of this study was to evaluate the relationship between cysteine-rich secretory protein-3 (CRISP-3) and some of its single-nucleotide polymorphisms (SNPs) with post-thawing semen quality in stallions. DNA was obtained from 100 stallions, regions of interest were amplified by polymerase chain reaction and sequenced. Evaluated SNPs within the equine CRISP-3 gene were CRISP3c.+199A > G (SNP1), CRISP3c.+566C > A (SNP2), CRISP3c.+622G > A (SNP3) and CRISP3c.+716A > G (SNP4). CRISP-3 protein content in seminal plasma was determined by enzyme-linked immunosorbent assay. Semen from 30 stallions was cryopreserved and post-thaw motility, kinetics, abnormal morphology (AM), sperm vitality (SV) and membrane integrity (MI) were evaluated. Generalized linear models were fitted and means were compared using Tukey’s test. Correlation and regression analyses were performed. For SNP1 and SNP3, the AA genotype had the highest results for motility and MI; for SNP2, the best results for motility and AM were obtained with the CC genotype. For SNP4, the GG genotype had the lowest results, except for MI. A high level of CRISP-3 protein in seminal plasma had the best results for motility, kinetics, SV and AM. In conclusion, there was a relationship between CRISP-3 genotype and seminal plasma protein and post-thawing semen quality in stallions.
Additional keywords: cryotolerance, freezing, seminal plasma, sequencing, sperm.
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