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Effect of immunisation against gonadotropin releasing hormone isoforms (mammalian GnRH-I, chicken gnRH-II and lamprey gnRH-III) on murine spermatogenesis

Khan, M.A.H. and Prevost, M. and Waterston, M.M. and Harvey, M.J. and Ferro, V.A. (2007) Effect of immunisation against gonadotropin releasing hormone isoforms (mammalian GnRH-I, chicken gnRH-II and lamprey gnRH-III) on murine spermatogenesis. Vaccine, 25 (11). pp. 2051-2063. ISSN 0264-410X

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Abstract

In mammals, the hypothalamic decapeptide, gonadotrophin releasing hormone (GnRH-I), is regarded as the major fertility regulating peptide. However, a range of isoforms also exists, varying only in the core region between amino acids 5–8. The physiological role of two of these, GnRH-II and GnRH-III, remains controversial, particularly with regard to fertility. The basis of the present study was to examine whether there is potential for GnRH-II and GnRH-III to be developed into highly specific vaccines, and to determine what the impact of their neutralisation would be on fertility. Computer modelling was used to predict how many common amino acids could be sequentially removed from the N-terminus, without loss of conformational structure. Sequences predicted to retain structure, were synthesised and conjugated to tetanus toxoid. Male mice were actively immunised, in study weeks 0, 2, 4 and 6 and peptide specific ELISA carried out. Mice immunised with TT-GnRH-I, TT-GnRH-II and TT-GnRH-III conjugates induced high antibody titres to the respective peptide. However, serum from TT-GnRH-I treated mice showed cross-reactivity to GnRH-II and GnRH-III peptides, and serum from TT-GnRH-II immunised mice showed cross-reactivity to GnRH-III. On the other hand, serum from only two of the TT-GnRH-III treated animals showed cross-reactivity to GnRH-II. Histological examination of the testes enabled comparative quantification of the disruption to spermatogenesis. Immunisation against TT-GnRH-I and TT-GnRH-III caused 66% and 68%, respectively, of seminiferous tubules viewed to show evidence of spermatogenesis, compared with 82% and 92% against TT-GnRH-II and untreated controls, respectively. Endocrine analysis revealed that only the TT-GnRH-I immunised animals showed significant reduction (p < 0.05) in follicle stimulating hormone, while testosterone levels were reduced in the TT-GnRH-I and TT-GnRH-III treated animals. Taken together, our data suggests that GnRH-I and GnRH-III are implicated in spermatogenesis, unlike GnRH-II.