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An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm

Bart Leemans (UGent) , Bart Gadella, Tom Stout, Hilde Nelis (UGent) , Maarten Hoogewijs (UGent) and Ann Van Soom (UGent)
(2015) REPRODUCTION. 150(3). p.193-208
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Abstract
Induction of hyperactivated motility is considered essential for triggering the release of oviduct-bound mammalian spermatozoa in preparation for fertilization. In this study, oviduct-bound stallion spermatozoa were exposed for 2 h to: i) pre-ovulatory and ii) post-ovulatory oviductal fluid; iii) 100% and iv) 10% follicular fluid (FF); v) cumulus cells, vi) mature equine oocytes, vii) capacitating and viii) non-capacitating medium. None of these triggered sperm release or hyperactivated motility. Interestingly, native FF was detrimental to sperm viability, an effect that was negated by heat inactivation, charcoal treatment and 30 kDa filtration alone or in combination. Moreover, sperm suspensions exposed to treated FF at pH 7.9 but not pH 7.4 showed Ca2+-dependent hypermotility. Fluo-4 AM staining of sperm showed elevated cytoplasmic Ca2+ in hyperactivated stallion spermatozoa exposed to treated FF at pH 7.9 compared to a modest response in defined capacitating conditions at pH 7.9 and no response in treated FF at pH 7.4. Moreover, 1 h incubation in alkaline, treated FF induced protein tyrosine phosphorylation in 20% of spermatozoa. None of the conditions tested induced widespread release of sperm pre-bound to oviduct epithelium. However, the hyperactivating conditions did induce release of 70-120 spermatozoa per oviduct explant, of which 48% showed protein tyrosine phosphorylation and all were acrosome-intact, but capable of acrosomal exocytosis in response to calcium ionophore. We conclude that, in the presence of elevated pH and extracellular Ca2+, a heat-resistant, hydrophilic, <30 kDa component of FF can trigger protein tyrosine phosphorylation, elevated cytoplasmic Ca2+ and hyperactivated motility in stallion sperm, but infrequent release of sperm pre-bound to oviduct epithelium.
Keywords
FEMALE REPRODUCTIVE-TRACT, PROTEIN-TYROSINE PHOSPHORYLATION, IN-VITRO PRODUCTION, BOVINE SPERM, HYPERACTIVATED MOTILITY, INTRACELLULAR CALCIUM, HUMAN SPERMATOZOA, POSTNATAL-DEVELOPMENT, HEAT INACTIVATION, BINDING PROTEINS

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MLA
Leemans, Bart et al. “An Alkaline Follicular Fluid Fraction Induces Capacitation and Limited Release of Oviduct Epithelium-bound Stallion Sperm.” REPRODUCTION 150.3 (2015): 193–208. Print.
APA
Leemans, B., Gadella, B., Stout, T., Nelis, H., Hoogewijs, M., & Van Soom, A. (2015). An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm. REPRODUCTION, 150(3), 193–208.
Chicago author-date
Leemans, Bart, Bart Gadella, Tom Stout, Hilde Nelis, Maarten Hoogewijs, and Ann Van Soom. 2015. “An Alkaline Follicular Fluid Fraction Induces Capacitation and Limited Release of Oviduct Epithelium-bound Stallion Sperm.” Reproduction 150 (3): 193–208.
Chicago author-date (all authors)
Leemans, Bart, Bart Gadella, Tom Stout, Hilde Nelis, Maarten Hoogewijs, and Ann Van Soom. 2015. “An Alkaline Follicular Fluid Fraction Induces Capacitation and Limited Release of Oviduct Epithelium-bound Stallion Sperm.” Reproduction 150 (3): 193–208.
Vancouver
1.
Leemans B, Gadella B, Stout T, Nelis H, Hoogewijs M, Van Soom A. An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm. REPRODUCTION. 2015;150(3):193–208.
IEEE
[1]
B. Leemans, B. Gadella, T. Stout, H. Nelis, M. Hoogewijs, and A. Van Soom, “An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm,” REPRODUCTION, vol. 150, no. 3, pp. 193–208, 2015.
@article{6940026,
  abstract     = {Induction of hyperactivated motility is considered essential for triggering the release of oviduct-bound mammalian spermatozoa in preparation for fertilization. In this study, oviduct-bound stallion spermatozoa were exposed for 2 h to: i) pre-ovulatory and ii) post-ovulatory oviductal fluid; iii) 100% and iv) 10% follicular fluid (FF); v) cumulus cells, vi) mature equine oocytes, vii) capacitating and viii) non-capacitating medium. None of these triggered sperm release or hyperactivated motility. Interestingly, native FF was detrimental to sperm viability, an effect that was negated by heat inactivation, charcoal treatment and 30 kDa filtration alone or in combination. Moreover, sperm suspensions exposed to treated FF at pH 7.9 but not pH 7.4 showed Ca2+-dependent hypermotility. Fluo-4 AM staining of sperm showed elevated cytoplasmic Ca2+ in hyperactivated stallion spermatozoa exposed to treated FF at pH 7.9 compared to a modest response in defined capacitating conditions at pH 7.9 and no response in treated FF at pH 7.4. Moreover, 1 h incubation in alkaline, treated FF induced protein tyrosine phosphorylation in 20% of spermatozoa. None of the conditions tested induced widespread release of sperm pre-bound to oviduct epithelium. However, the hyperactivating conditions did induce release of 70-120 spermatozoa per oviduct explant, of which 48% showed protein tyrosine phosphorylation and all were acrosome-intact, but capable of acrosomal exocytosis in response to calcium ionophore. We conclude that, in the presence of elevated pH and extracellular Ca2+, a heat-resistant, hydrophilic, <30 kDa component of FF can trigger protein tyrosine phosphorylation, elevated cytoplasmic Ca2+ and hyperactivated motility in stallion sperm, but infrequent release of sperm pre-bound to oviduct epithelium.},
  author       = {Leemans, Bart and Gadella, Bart and Stout, Tom and Nelis, Hilde and Hoogewijs, Maarten and Van Soom, Ann},
  issn         = {1470-1626},
  journal      = {REPRODUCTION},
  keywords     = {FEMALE REPRODUCTIVE-TRACT,PROTEIN-TYROSINE PHOSPHORYLATION,IN-VITRO PRODUCTION,BOVINE SPERM,HYPERACTIVATED MOTILITY,INTRACELLULAR CALCIUM,HUMAN SPERMATOZOA,POSTNATAL-DEVELOPMENT,HEAT INACTIVATION,BINDING PROTEINS},
  language     = {eng},
  number       = {3},
  pages        = {193--208},
  title        = {An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm},
  url          = {http://dx.doi.org/10.1530/REP-15-0178},
  volume       = {150},
  year         = {2015},
}

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