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Hormone-Induced Sperm-Release in the Critically University of Wollongong Research Online Faculty of Science, Medicine and Health - Papers: Part B Faculty of Science, Medicine and Health 1-1-2019 Hormone-induced sperm-release in the critically endangered Booroolong frog (Litoria booroolongensis): Effects of gonadotropin-releasing hormone and human chorionic gonadotropin Aimee J. Silla University of Wollongong, [email protected] Michael McFadden University of Wollongong, [email protected] Phillip G. Byrne University of Wollongong, [email protected] Follow this and additional works at: https://ro.uow.edu.au/smhpapers1 Publication Details Citation Silla, A. J., McFadden, M., & Byrne, P. G. (2019). Hormone-induced sperm-release in the critically endangered Booroolong frog (Litoria booroolongensis): Effects of gonadotropin-releasing hormone and human chorionic gonadotropin. Faculty of Science, Medicine and Health - Papers: Part B. Retrieved from https://ro.uow.edu.au/smhpapers1/579 Research Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library: [email protected] Hormone-induced sperm-release in the critically endangered Booroolong frog (Litoria booroolongensis): Effects of gonadotropin-releasing hormone and human chorionic gonadotropin Abstract Research into the development of reproductive technologies for amphibians has increased in recent years due to the rapid decline of amphibian species globally. Reproductive technologies have great potential to overcome captive breeding failure and improve the propagation and genetic management of threatened species. However, the incorporation of these technologies into conservation breeding programs has been protracted, primarily as a result of trial-and-error approaches to the refinement of hormone therapies. The present study investigated the effects of: (1) GnRH-a dose (0, 0.5, 1, 2, 4, 8 or 16 μg g -1 ), and (2) hCG dose (0, 2.5, 5, 10, 20 or 40 IU g -1 ), on the sperm-release response of the critically endangered Booroolong frog. Administration of GnRH-a at a dose of 0.5 μg g -1 resulted in the greatest number of sperm released (mean total sperm = 3.5 x10 6, n = 11). Overall, hCG was more effective at eliciting spermiation in Booroolong frogs, with peak sperm release (mean total sperm = 25.1 x10 6, n = 10) occurring in response to a dose of 40 IU g -1 . Sperm output in response to 40 IU g -1 hCG was greatest between 1 and 6 h and steadily declined between 8 and 24 h post-hormone administration. Percent sperm motility peaked between 4 and 10 h (58.1-62.7%), and sperm velocity between 4 and 12 h (24.3-27.2 μm s -1 ). Booroolong frogs join a small, but growing number of amphibian species that exhibit improved spermiation in response to hCG. Further research is required to identify optimal hormone-induction protocols for threatened amphibians and expedite the incorporation of reproductive technologies into CBPs. Publication Details Silla, A. J., McFadden, M. S. & Byrne, P. G. (2019). Hormone-induced sperm-release in the critically endangered Booroolong frog (Litoria booroolongensis): Effects of gonadotropin-releasing hormone and human chorionic gonadotropin. Conservation Physiology, 7 (1), 1-10. This journal article is available at Research Online: https://ro.uow.edu.au/smhpapers1/579 Volume 7 • 2019 10.1093/conphys/coy080 Research article Hormone-induced sperm-release in the critically Downloaded from https://academic.oup.com/conphys/article-abstract/7/1/coy080/5316805 by University of Wollongong user on 08 March 2019 endangered Booroolong frog (Litoria booroolongensis): effects of gonadotropin- releasing hormone and human chorionic gonadotropin Aimee J. Silla1,*, Michael S. McFadden2 and Phillip G. Byrne1 1School of Earth, Atmospheric and Life Sciences, University of Wollongong, Wollongong, NSW 2522, Australia 2Herpetofauna Department, Taronga Conservation Society Australia, PO Box 20, Mosman, NSW 2088, Australia *Corresponding author: Email: [email protected] .............................................................................................................................................................. Research into the development of reproductive technologies for amphibians has increased in recent years due to the rapid decline of amphibian species globally. Reproductive technologies have great potential to overcome captive breeding fail- ure and improve the propagation and genetic management of threatened species. However, the incorporation of these technologies into conservation breeding programs has been protracted, primarily as a result of trial-and-error approaches to the refinement of hormone therapies. The present study investigated the effects of: (1) GnRH-a dose (0, 0.5, 1, 2, 4, 8 or 16 μgg−1), and (2) hCG dose (0, 2.5, 5, 10, 20 or 40 IU g−1), on the sperm-release response of the critically endangered Booroolong frog. Administration of GnRH-a at a dose of 0.5 μgg−1 resulted in the greatest number of sperm released (mean total sperm = 3.5 ×106, n = 11). Overall, hCG was more effective at eliciting spermiation in Booroolong frogs, with peak sperm release (mean total sperm = 25.1 ×106, n = 10) occurring in response to a dose of 40 IU g−1. Sperm output in response to 40 IU g−1 hCG was greatest between 1 and 6 h and steadily declined between 8 and 24 h post-hormone admin- istration. Percent sperm motility peaked between 4 and 10 h (58.1–62.7%), and sperm velocity between 4 and 12 h (24.3–27.2 μms−1). Booroolong frogs join a small, but growing number of amphibian species that exhibit improved sper- miation in response to hCG. Further research is required to identify optimal hormone-induction protocols for threatened amphibians and expedite the incorporation of reproductive technologies into CBPs. Key words: Amphibian, captive breeding, conservation, gamete release, reproduction, reproductive technologies, spermiation Editor: Dr Steven Cooke Received 18 September 2018; Revised 6 December 2018; Editorial Decision 16 December 2018; accepted 3 January 2019 Cite as: Silla AJ, McFadden MS, Byrne PG (2019) Hormone-induced sperm-release in the critically endangered Booroolong frog (Litoria booroolongensis): effects of gonadotropin-releasing hormone and human chorionic gonadotropin. Conserv Physiol 7(1): coy080; doi:10.1093/ conphys/coy080. .............................................................................................................................................................. .............................................................................................................................................................. © The Author(s) 2019. Published by Oxford University Press and the Society for Experimental Biology. 1 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Research article Conservation Physiology • Volume 7 2019 .............................................................................................................................................................. Introduction respond more favourably to hCG, including several species from the Bufonid, Limnodynastid and Pelodryadid families Ongoing species decline and extinction continues to threaten (Kouba et al.,2012; Clulow et al.,2018; Silla and Roberts, vertebrate biodiversity across all taxonomic groups and geo- 2012). The inconsistent response to hCG administration is graphic locations globally (Ceballos et al., 2015). Among thought to be driven by interspecific differences in LH-receptor vertebrates, amphibians have experienced an exceptionally affinities (Clulow et al.,2014), which may reflect the divergent rapid loss of biodiversity, with at least 2100 species at immi- evolution of affinities of LH-receptors among amphibian fam- Downloaded from https://academic.oup.com/conphys/article-abstract/7/1/coy080/5316805 by University of Wollongong user on 08 March 2019 nent risk of extinction, representing 31.8% of all described ilies (Silla and Roberts, 2012). At present, insufficient data species (IUCN, 2017). In response to this extinction crisis, exists directly comparing the gamete-release response of conservation breeding programs (CBPs) have been estab- amphibians from different lineages to the administration of lished for a number of threatened amphibian species with GnRH-a and hCG. Consequently, our capacity to predict the some evidence of successful reintroduction and species recov- response of novel species is poor, and hormonal induction pro- ery (Harding et al., 2016). Despite such celebrated triumphs, tocols need to be developed on a species-specific basis. disparity remains between the scale of the global amphibian crisis and the response from the conservation community, Central to the development of hormonal induction proto- cols for novel species is the establishment of dose-response with current estimates indicating that a mere 2.9% (213 spe- fi cies) of all amphibian species have been the subject of CBPs curves for each hormone type (GnRH-a and hCG), and veri - (Harding et al., 2016). One of the major challenges hinder- cation of optimal gamete-collection times post-administration (PA). If suboptimal or supraoptimal hormone doses are admi- ing the success of CBPs, and limiting the establishment of nistered, gamete release will not occur or gametes will be new programs, is captive breeding failure.
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