Viable and morphologically normal boar spermatozoa alter the expression of heat-shock protein genes in oviductal epithelial cells during co-culture in vitro
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The principal aim of this study was to determine if boar spermatozoa influence the expression of four selected chaperone and heat-shock protein (HSP) genes-namely clusterin (CLU), HSP90AA1, HSPA5, and HSPA8-in oviductal epithelial cells (OECs) during in vitro co-culture. All corresponding proteins of these genes were previously identified in a sperm-interacting, 70-kDa soluble fraction derived from apical plasma membranes of OECs. The present study also sought to determine whether or not: (i) spermatozoa must directly bind to OEC for an effect on gene expression to be elicited and (ii) reproductive and nonreproductive epithelial cell types (LLC-PK1, pig kidney) respond equivalently, in terms of alterations in chaperone and HSP gene expression, during co-culture with sperm. Spermatozoa induced a significant upregulation (P < 0.05) in HSP90AA1 and HSPA5 in OECs after 3 hr, and in HSPA8 after 6 hr of co-culture when they were in direct contact with epithelial cells. Conversely, no upregulation of HSP transcription was observed when spermatozoa did not directly bind to OECs. Spermatozoa also induced a significant upregulation (P < 0.05) of the same three genes when in direct contact with LLC-PK1 cells, but the timing occurred later than with OECs. Interestingly, the extent of HSP gene upregulation induced by direct contact of spermatozoa with epithelial cells was dependent on sperm-binding index and on the viability and morphological quality of the bound sperm population. In conclusion, the upregulation of HSP genes caused by direct contact between spermatozoa and OECs, rather than nonreproductive epithelial cells, suggests HSPs could play an integral role in the modulation of sperm function in the oviductal reservoir.
|Journal||Molecular Reproduction and Development|
|Early online date||4 Aug 2014|
|Publication status||Published - Sep 2014|
Rights statement: This is the peer reviewed version of the following article: Yeste, M., Holt, W. V., Bonet, S., Rodríguez-Gil, J. E. and Lloyd, R. E. (2014), Viable and morphologically normal boar spermatozoa alter the expression of heat-shock protein genes in oviductal epithelial cells during co-culture in vitro. Mol. Reprod. Dev., 81: 805–819, which has been published in final form at 10.1002/mrd.22350. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Accepted author manuscript (Post-print), 196 KB, PDF document