Two-dimensional polyacrylamide gel electrophoresis of equine seminal plasma proteins and their relation with semen freezability

Cysteine-rich secretory protein-3 (CRISP-3) and some of its nonsynonymous polymorphism have been related to the fertility and freezability of stallion semen; however, the role of the CRISP-3 gene and its seminal plasma protein in the raw semen quality is still unknown. The aim of this study was to evaluate the relationship of CRISP-3 with semen quality in stallions. DNA was obtained from blood samples of 100 stallions, from which 30 stallions were randomly selected to obtain 60 ejaculates. Through PCR amplification and sequencing, the variation of four nonsynonymous SNPs from CRISP-3 was identified and haplotypes were derived. Semen quality was assessed through the total motility (MOT), sperm vitality (SV), normal morphology (NM), functional integrity of membrane (MI) and a seminal quality index (SQi). CRISP-3 protein content of seminal plasma (SP) was determined by ELISA. The effect of the genotype, the haplotype and the concentration of the CRISP-3 protein on the seminal quality were evaluated through generalized linear models and linear regression analyses. Homozygous genotypes for SNP1, SNP2 and SNP3 and the heterozygous genotype for SNP4 showed a positive effect on seminal quality. Different haplotypes with positive effect on MOT, SV, NM, MI and SQi were identified. The allelic substitution analysis resulted in positive regression coefficients for MOT (SNP2) and MI (SNP2 and SNP3). A high level of CRISP-3 resulted in a higher MOT and SQi. It is concluded that the quality of stallion semen is influenced by the genotype of CRISP-3 and the concentration of CRISP-3 protein in SP.

Section: Discussionmentioning

confidence: 99%

Relationship of cysteine‐rich secretory protein‐3 gene and protein with semen quality in stallions

Betancur

Rojano

Suarez

2018

“…2004). Interestingly, proteins of this heterodimer have been both negatively (HSP1) and positively (HSP2) correlated with the ability of stallion spermatozoa to survive freezing (Jobim et al. 2011), and the functional significance of this effect is unclear.…”

Section: Reasons For Species Differences In the Function Of Bsps And mentioning

confidence: 99%

Seminal Plasma and its Effect on Ruminant Spermatozoa During Processing

Leahy

Graaf

2012

Contents Seminal plasma can both inhibit and stimulate sperm function, making its use as a supportive medium somewhat contradictory. These effects are directed by the multifunctional action of numerous inorganic and organic components, but it is the direct association of seminal plasma proteins with the sperm membrane that is thought to exert the most significant response. In vitro handling of spermatozoa in preparation for artificial insemination may involve washing, dilution, cooling, freezing, re‐warming and sex‐sorting. These processes can alter proteins of the sperm surface and reduce seminal plasma in the sperm environment. This, among other factors, may destabilize the sperm membrane and reduce the fertilizable lifespan of spermatozoa. Such handling‐induced damage may be prevented or reversed through supplementation of seminal plasma, but the effectiveness of this technique differs with species, and the source and subsequent treatment of both spermatozoa and seminal plasma. Seminal plasma appears to act as a protective medium during in vitro processing of ram spermatozoa, but this does not appear to be the case for bull spermatozoa. The reasons for this divergent effect will be discussed with particular emphasis on the influence of the major proteins of ruminant seminal plasma, known as BSP proteins. The biochemical and biophysical properties of these proteins are well documented, and this information has provided greater insight into the signalling pathways of capacitation and the protective action of extender components.

“…In some species, seminal plasma proteins related to fertility (Killian et al 1993;Brandon et al 1999;Ashrafzadeh et al 2013), cold-shock damage sperm membrane resistance (Barrios et al 2000;P erez-P e et al 2001), sperm tolerance to freezing (Goularte et al 2014), freezability (Jobim et al 2011) and semen quality (Cardozo et al 2006;. De Souza et al 2007;Arruda-Alencar et al 2012;Nandre et al 2013;Kiso et al 2013) have been identified.…”

Section: Introductionmentioning

confidence: 99%

Can the Genetic Origin Affect Rabbit Seminal Plasma Protein Profile along the Year?

Casares-Crespo

Talaván

Viudes-de-Castro

2016

The study was designed to evaluate the influence of genetic origin on rabbit seminal plasma protein profile variation along the year. Seminal plasma of rabbits from line A (maternal line) and R (paternal line) collected during a natural year was subjected to polyacrylamide gel electrophoresis (SDS-PAGE). The electrophoretic profile of rabbit seminal plasma resulted in multiple protein bands of different intensity ranging from 9 to 240 kDa. Results showed that seven protein bands were significantly different between genetic lines, and among these, three protein bands were significantly different between seasons. The differentially expressed proteins were identified by MALDI-TOF/TOF or LC-MS/MS analysis and were the following ones: FAM115E-like (220, 113 and 59 kDa), ectonucleoside triphosphate diphosphohydrolase 3 isoform X2 (72 kDa), annexin A5 (32 kDa), lipocalin allergen Ory c 4 precursor (19 kDa), and haemoglobin subunit zetalike (13 kDa) between genetic lines and FAM115E-like (113 kDa), haemoglobin subunit zetalike (13 kDa) and β-nerve growth factor (12 kDa) between seasons. These results indicate that proteins from rabbit seminal plasma are under both seasonal control and genetic control. Furthermore, the differential presence of these proteins could be one of the causes explaining the differences observed in fertility and seminal parameters between these two lines in earlier studies.