Effect of porcine zonae pellucidae immunisation on ovarian follicular development and endocrine function in domestic ewes (Ovis aries)
M. A. Stoops A F , I. K. M. Liu B , S. E. Shideler C , B. L. Lasley C , R. A. Fayrer-Hosken D , K. Benirschke E , K. Murata A , E. M. G. van Leeuwen B and G. B. Anderson AA Department of Animal Science, University of California, Davis, CA 95616, USA.
B Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
C Center for Health and the Environment, University of California, Davis, CA 95616, USA.
D Department of Large Animal Medicine, Physiology and Pharmacology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602, USA.
E University of California, San Diego, Medical Center, San Diego, CA 92103, USA.
F Corresponding author. Email: monica.stoops@cincinnatizoo.org
Reproduction, Fertility and Development 18(6) 667-676 https://doi.org/10.1071/RD05045
Submitted: 14 April 2005 Accepted: 11 April 2006 Published: 22 June 2006
Abstract
Domestic ewes (Ovis aries) were immunised with porcine zonae pellucidae (pZP) or pZP conjugated to keyhole limpet haemocyanin (KLH) in adjuvant(s) to examine the feasibility of the species to serve as a model for further development of pZP-based vaccines in ungulates. Two immunisation groups were employed, with a third group receiving only adjuvant (n = 5 per group). Early in the study, oestrous activity was monitored by the use of a vasectomised ram fitted with a marking harness. Eventually, ewes were exposed to an intact ram for breeding. In addition, weekly serum and every-other-day faecal samples were collected to measure pZP antibodies and progesterone metabolite concentrations respectively. At the conclusion of the study, fecundity was established, and ovarian tissue was examined. Ewes immunised against pZP : KLH with adjuvant produced minimal antibody absorbance levels, displayed normal oestrous cycles, became pregnant upon introduction of the intact ram and exhibited normal ovarian histopathology. Ewes immunised against pZP with adjuvant produced high antibody absorbance levels, were acyclic following primary immunisation and were infertile. Examination of the ovarian tissue revealed atrophic changes that included: (1) the absence of growing follicles; (2) significant reduction in the number of primordial follicles; and (3) the presence of abnormal granulosa cell clusters lacking oocytes. Antisera displayed immunoreactivity to the major components of pZP, and immunohistochemical labelling of ovarian tissue showed specificity to the ZP. These data are the first generated in an ungulate species showing deleterious effects of pZP immunisation on folliculogenesis and oestrous cyclicity.
Acknowledgments
This work was supported by funds from: the Department of Animal Science, University of California, Davis; National Institute of Health, Fertilization and Early Development Training Grant; US EPA (R825433), Center for Ecological Health Research, University of California, Davis. We thank Maria Morales for conducting the ELISA on sheep serum samples; Alice Moyer for assistance during handling and bleeding sheep; Tinyee Hoang, Sheri Looper, Brandi Macho, Beckye Stanton and Heather Todd for sheep faecal sample collection assistance; and Dana Van Liew for sheep husbandry.
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