Effects of oviductal proteins, including heat shock 70 kDa protein 8, on survival of ram spermatozoa over 48 h in vitro
R. E. Lloyd A D , R. M. A. Elliott A , A. Fazeli B , P. F. Watson C and W. V. Holt AA 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 Corresponding author. Email: rhiannon.lloyd@ioz.ac.uk
Reproduction, Fertility and Development 21(3) 408-418 https://doi.org/10.1071/RD08204
Submitted: 21 September 2008 Accepted: 20 October 2008 Published: 4 March 2009
Abstract
Following insemination, ram spermatozoa are transported to the isthmus region of the oviduct where they bind to the oviductal epithelial cells (OEC), remaining viable for several hours. The aim of the present study was to begin to decipher which component(s) of the ewe oviduct actively participates in maintaining the viability of ram spermatozoa. A series of experiments was conducted to investigate whether: (1) soluble OEC apical plasma membrane proteins (sAPM) isolated from ewes prolong survival of ram spermatozoa over an extended (48 h) coincubation period at 39°C; (2) a recombinant form of one of these oviductal proteins, namely heat shock 70 kDa protein 8 (HSPA8), prolongs survival of ram spermatozoa; and (3) pretreatment with HSPA8 antibody compromises the ability of sAPM to prolong the survival of ram spermatozoa. Both sAPM and recombinant HSPA8 had a beneficial effect on the viability of ram spermatozoa during coincubation, although both these effects were dose dependent. In contrast, pretreatment with HSPA8 antibody significantly negated the ability of sAPM to maintain the viability of ram spermatozoa. These findings suggest that HSPA8 is an active component of the ewe oviduct that participates in maintaining the viability of ram spermatozoa. This is a potentially valuable observation given that there is a great deal of room for improving existing diluents for storing fresh ram semen.
Acknowledgement
The authors are grateful to Defra (London, UK), Innovis Ltd (Aberystwyth, UK) and IMV Technologies (L’Aigle Cedex, France) for funding this study. The authors also thank Mrs Christine Bruce for preparing the oviductal sections for histological analysis.
Arispe, N. , Doh, M. , Simakova, O. , Kurganov, B. , and De Maio, A. (2004). Hsc70 and Hsp70 interact with phosphatidylserine on the surface of PC12 cells resulting in a decrease of viability. FASEB J. 18, 1636–1645.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Fazeli, A. , Elliott, R. M. , Duncan, A. E. , Moore, A. , Watson, P. F. , and Holt, W. V. (2003). In vitro maintenance of boar sperm viability by a soluble fraction obtained from oviductal apical plasma membrane preparations. Reproduction 125, 509–517.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Meister, A. , Griffith, O. W. , Novogrodsky, A. , and Tate, S. S. (1979). New aspects of glutathione metabolism and translocation in mammals. Ciba Found. Symp. 72, 135–161.
| PubMed |
Olson, G. E. , Winfrey, V. P. , Nagdas, S. K. , and Melner, M. H. (2004). Region-specific expression and secretion of the fibrinogen-related protein, fgl2, by epithelial cells of the hamster epididymis and its role in disposal of defective spermatozoa. J. Biol. Chem. 279, 51 266–51 274.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Pouliot, J. F. , and Beliveau, R. (1994). Extraction of brain capillary membrane proteins using Triton X-114. Biochem. Mol. Biol. Int. 34, 1075–1084.
| PubMed | CAS |
Salamon, S. , and Maxwell, W. M. (2000). Storage of ram semen. Anim. Reprod. Sci. 62, 77–111.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Satake, N. , Elliott, R. M. , Watson, P. F. , and Holt, W. V. (2006). Sperm selection and competition in pigs may be mediated by the differential motility activation and suppression of sperm subpopulations within the oviduct. J. Exp. Biol. 209, 1560–1572.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Smith, T. T. , and Nothnick, W. B. (1997). Role of direct contact between spermatozoa and oviductal epithelial cells in maintaining rabbit sperm viability. Biol. Reprod. 56, 83–89.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Trubner, M. , Glander, H. J. , and Schaller, J. (1997). Localization of adhesion molecules on human spermatozoa by fluorescence microscopy. Andrologia 29, 253–260.
| PubMed | CAS |
