Novel insights into the role of cell-free seminal mRNAs on semen quality and cryotolerance of spermatozoa in bulls (Bos taurus)
Munivenkatappa Shilpa A B , Sellappan Selvaraju A C , Venkataswamy GirishKumar B , Sivashanmugam Parthipan A , Krishnan B. Binsila A , Arunachalam Arangasamy A and Janivara Parameshwaraiah Ravindra A
+ Author Affiliations
- Author Affiliations
A Reproductive Physiology Laboratory, Animal Physiology Division, Indian Council of Agricultural Research – National Institute of Animal Nutrition and Physiology, Adugodi, Bengaluru 560030, India.
B Department of Veterinary Biochemistry, Veterinary College, Karnataka Veterinary and Fisheries Sciences University, Bengaluru 560024, India.
C Corresponding author. Email: selvaraju.s@icar.gov.in
Reproduction, Fertility and Development 29(12) 2446-2456 https://doi.org/10.1071/RD16290
Submitted: 25 January 2016 Accepted: 9 May 2017 Published: 14 June 2017
Abstract
The aim of the present study was to ascertain the effectiveness of seminal plasma mRNAs as markers to assess the reproductive performance of bulls. Semen samples (33 ejaculates) from 11 bulls were evaluated for sperm kinematic and functional parameters. Total RNA was isolated from cell-free seminal (cfs) using TRIzol LS reagent and the concentration of cfs-RNA was 24.4 ± 2.3 µg mL−1 seminal plasma. The cfs-RNA was fragmented to a size of 25–500 bp. Of the cfs-mRNAs screened using real time PCR, expression of protamine 1 (PRM1) was positively (P < 0.05) associated with the mitochondrial membrane potential of raw semen, whereas expression of Fas Ligand (FASLG) was negatively (P < 0.05) associated with sperm velocity, membrane integrity and chromatin distribution in post-thaw semen samples. The percentage of Type A spermatozoa (amplitude of lateral movement of head >2.5 μm and straightness >85%) in raw semen was positively (P < 0.05) associated with bone morphogenetic protein 2 (BMP2), ubiquitin conjugating enzyme E2D3 (UBE2D3), tumour-associated necrotic factor-associated death domain (TRADD) and caspase-3 (CASP3) expression. Nerve growth factor (NGF) expression was positively (P < 0.05) associated with the maintenance of post-thaw functional membrane integrity in spermatozoa and could be used to assess the cryotolerance of bull semen. In conclusion, the expression of cfs mRNAs can be used to assess the reproductive performance of males and to predict the sensitivity of spermatozoa to cryoinjury.
Additional keywords: cryoinjury, transcript expression.
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