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Vertebrate reproductive science and technology
RESEARCH ARTICLE

The oviducal protein, heat-shock 70-kDa protein 8, improves the long-term survival of ram spermatozoa during storage at 17°C in a commercial extender

R. E. Lloyd A D E , A. Fazeli B , P. F. Watson C and W. V. Holt A
+ Author Affiliations
- Author Affiliations

A Institute of Zoology, Regent’s Park, London NW1 4RY, UK.

B Academic Unit of Reproductive and Developmental Medicine, The University of Sheffield, Level 4, The Jessop Wing, Tree Root Walk, Sheffield S10 2SF, UK.

C Royal Veterinary College, Royal College Street, London, NW1 0TU, UK.

D Present address: Nuffield Building, Institute of Zoology, Regent’s Park, London NW1 4RY, UK.

E Corresponding author. Email: rhiannon.lloyd@ioz.ac.uk

Reproduction, Fertility and Development 24(4) 543-549 https://doi.org/10.1071/RD11173
Submitted: 6 June 2011  Accepted: 14 September 2011   Published: 16 November 2011

Abstract

Poor fertility rates are often observed when fresh ram semen stored in conventional extenders is used for cervical artificial insemination (AI). Heat-shock 70-kDa protein 8 (HSPA8), found within the oviduct, prolongs boar, ram and bull sperm survival at body temperatures in vitro. Here, we aimed to determine whether supplementing extenders (INRA-96 and RSD-1) with HSPA8 (4 µg mL–1) would improve their performance in maintaining freshly collected ram sperm viability and sperm nuclear DNA integrity during storage over 48 h at 17°C. Sperm function was assessed at 1, 6, 24 and 48 h and this experiment was repeated using 25 × 106 and 800 × 106 spermatozoa mL–1. INRA96 supplemented with HSPA8 maintained sperm viability significantly better than INRA96 alone at both sperm concentrations. However, sperm nuclear DNA fragmentation (DF) increased significantly during storage using the higher sperm concentration, irrespective of the extender and the protein treatment used. Increasing levels of sperm nuclear DF over time could explain why poor fertility rates are often observed following cervical AI using stored ram semen. However, further research is required to ascertain whether supplementing the commercially available INRA96 extender with HSPA8 will improve fertility rates following cervical AI using stored ram semen.

Additional keywords: DNA fragmentation, fresh sperm, HSPA8, liquid-storage.


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