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Reproduction (2007) 133 909-917
DOI: 10.1530/REP-06-0228
Copyright © 2007 Society for Reproduction and Fertility
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RESEARCH

Detection of osteopontin on Holstein bull spermatozoa, in cauda epididymal fluid and testis homogenates, and its potential role in bovine fertilization

David W Erikson, Amy L Way1, David A Chapman and Gary J Killian

Department of Dairy & Animal Science, John O Almquist Research Center, The Pennsylvania State University, University Park, Pennsylvania 16802, USA and 1 Department of Health Science, Lock Haven University, Clearfield Campus, Clearfield, Pennsylvania 16830, USA

Correspondence should be addressed to G J Killian; Email: gkillian{at}psu.edu

Osteopontin (OPN) is a secreted extracellular matrix phosphoprotein identified in various tissues and fluids including those of the male and female reproductive tracts. OPN was previously identified as a 55 kDa high fertility marker in Holstein bull seminal plasma, produced by the ampulla and the vesicular gland. The objectives of this study were to characterize OPN on ejaculated and cauda epididymal sperm using immunofluorescence and western blot analysis, and to assess the role of sperm OPN in fertilization. Solubilized sperm membrane proteins from ejaculated and cauda epididymal sperm were separated by 1D SDS-PAGE, transferred to nitrocellulose, and probed with an antibody to bovine milk OPN. A 35 kDa protein was detected by this antibody in both ejaculated and cauda epididymal sperm membranes. Analyses also recognized OPN at 55 and 25 kDa in cauda epididymal fluid and testicular parenchyma homogenates respectively. Immunofluorescent analysis of ejaculated and cauda epididymal sperm showed OPN localization in a well-defined band in the postacrosomal region of the sperm head and also on the midpiece. Results of in vitro fertilization experiments showed that sperm treated with an antibody to OPN fertilized fewer oocytes than sperm treated with control medium while increasing incidence of polyspermy, suggesting a role of sperm-associated OPN in fertilization and a block to polyspermy. These studies demonstrate that OPN exists at multiple molecular weight forms in the bull reproductive tract and its presence on ejaculated sperm may signal its importance in fertilization by interacting with integrins or other proteins on the oocyte plasma membrane.




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E. L. Karcher, D. O. Bayles, J. P. Bannantine, D. C. Beitz, and J. R. Stabel
Osteopontin: A Novel Cytokine Involved in the Regulation of Mycobacterium avium subspecies paratuberculosis Infection in Periparturient Dairy Cattle
J Dairy Sci, August 1, 2008; 91(8): 3079 - 3091.
[Abstract] [Full Text] [PDF]


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