Sperm parameters following hormonal induction of spermiation in an endangered frog [the alpine tree frog] (Litoria verreauxii alpina)
Thien H. Pham A and Laura A. Brannelly
A *
+ Author Affiliations
- Author Affiliations
A Faculty of Veterinary and Agricultural Sciences, Melbourne Veterinary School, The University of Melbourne, Werribee, Vic. 3030, Australia.
Reproduction, Fertility and Development 34(13) 867-874 https://doi.org/10.1071/RD22009
Published online: 27 May 2022
© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing. This is an open access article distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND)
Abstract
Context: With global amphibian biodiversity rapidly declining, improving reproductive technology outcomes has become essential. Captive breeding programs have struggled because amphibian breeding physiology often requires specific environmental cues that reproductive technologies can circumvent.
Aims: This study tests the efficiency of hormonal induction by evaluating sperm quality in the endangered Litoria verreauxii alpina.
Methods: We assessed the effects of exogenous hormones – gonadotrophin-releasing hormone agonist (GnRH-a, Lucrin), and human chorionic gonadotrophin (hCG, Chorulon) – on sperm quality.
Key results: Hormone induction with hCG showed high efficacy while GnRH-a yielded a low response in producing sperm. Sperm quantity was affected by time post injection, with the greatest quantity at 1 h post injection. Sperm quality was also affected by time, where the sperm head size decreased by 11% at 7 h post injection.
Conclusions: Based on the results from this study, we recommend that that sperm be collected soon after induction, and not more than 4 h post induction in L. v. alpina. More work needs to be completed before recommending an optimal hormone induction method and dose, but 120 IU of hCG per male was successful for inducing spermiation.
Implications: This study represents a useful starting point for developing assisted reproductive techniques for non-model organisms.
Keywords: amphibian reproduction, artificial reproductive technologies, captive breeding, endangered species, hormonal induction, IVF, sperm viability, spermic urine.
References
Arregui, L, Diaz-Diaz, S, Alonso-López, E, and Kouba, AJ (2019). Hormonal induction of spermiation in a Eurasian bufonid (Epidalea calamita). Reproductive Biology and Endocrinology 17, 92.| Hormonal induction of spermiation in a Eurasian bufonid (Epidalea calamita).Crossref | GoogleScholarGoogle Scholar | 31711511PubMed |
Arregui, L, Bóveda, P, Gosálvez, J, and Kouba, AJ (2020). Effect of seasonality on hormonally induced sperm in Epidalea calamita (Amphibia, Anura, Bufonidae) and its refrigerated and cryopreservated storage. Aquaculture 529, 735677.
| Effect of seasonality on hormonally induced sperm in Epidalea calamita (Amphibia, Anura, Bufonidae) and its refrigerated and cryopreservated storage.Crossref | GoogleScholarGoogle Scholar |
Bates, D, Mächler, M, Bolker, B, and Walker, S (2015). Fitting linear mixed-effects models using lme4. Journal of Statistics Software 67, 1–48.
| Fitting linear mixed-effects models using lme4.Crossref | GoogleScholarGoogle Scholar |
Brannelly, LA, Hunter, DA, Lenger, D, Scheele, BC, Skerratt, LF, and Berger, L (2015). Dynamics of chytridiomycosis during the breeding season in an Australian alpine amphibian. PLoS ONE 10, e0143629.
| Dynamics of chytridiomycosis during the breeding season in an Australian alpine amphibian.Crossref | GoogleScholarGoogle Scholar | 26629993PubMed |
Brannelly, LA, Ohmer, MEB, and Richards-Zawacki, CL (2019). Artificial reproduction using leuprolide acetate in the frog Rana pipiens. Herpetological Journal 29, 125–130.
| Artificial reproduction using leuprolide acetate in the frog Rana pipiens.Crossref | GoogleScholarGoogle Scholar |
Browne, RK, Seratt, J, Vance, C, and Kouba, A (2006). Hormonal priming, induction of ovulation and in-vitro fertilization of the endangered Wyoming toad (Bufo baxteri). Reproductive Biology and Endocrinology 4, 34.
| Hormonal priming, induction of ovulation and in-vitro fertilization of the endangered Wyoming toad (Bufo baxteri).Crossref | GoogleScholarGoogle Scholar | 16790071PubMed |
Browne, RK, Kaurova, SA, Uteshev, VK, Shishova, NV, McGinnity, D, Figiel, CR, Mansour, N, Agnew, D, Wu, M, Gakhova, EN, Dzyuba, B, and Cosson, J (2015). Sperm motility of externally fertilizing fish and amphibians. Theriogenology 83, 1–13.e8.
| Sperm motility of externally fertilizing fish and amphibians.Crossref | GoogleScholarGoogle Scholar | 25442393PubMed |
Byrne, PG, and Silla, AJ (2010). Hormonal induction of gamete release, and in-vitro fertilisation, in the critically endangered Southern Corroboree Frog, Pseudophryne corroboree. Reproductive Biology and Endocrinology 8, 144.
| Hormonal induction of gamete release, and in-vitro fertilisation, in the critically endangered Southern Corroboree Frog, Pseudophryne corroboree.Crossref | GoogleScholarGoogle Scholar | 21114857PubMed |
Byrne, PG, Simmons, LW, and Roberts, JD (2003). Sperm competition and the evolution of gamete morphology in frogs. Proceedings of the Royal Society B: Biological Sciences 270, 2079–2086.
| Sperm competition and the evolution of gamete morphology in frogs.Crossref | GoogleScholarGoogle Scholar | 14561298PubMed |
Clulow, J, Pomering, M, Herbert, D, Upton, R, Calatayud, N, Clulow, S, Mahony, MJ, and Trudeau, VL (2018). Differential success in obtaining gametes between male and female Australian temperate frogs by hormonal induction: a review. General and Comparative Endocrinology 265, 141–148.
| Differential success in obtaining gametes between male and female Australian temperate frogs by hormonal induction: a review.Crossref | GoogleScholarGoogle Scholar | 29859744PubMed |
Collins, JP (2010). Amphibian decline and extinction: what we know and what we need to learn. Diseases of Aquatic Organisms 92, 93–99.
| Amphibian decline and extinction: what we know and what we need to learn.Crossref | GoogleScholarGoogle Scholar | 21268970PubMed |
Cornwallis, CK, and Birkhead, TR (2007). Changes in sperm quality and numbers in response to experimental manipulation of male social status and female attractiveness. The American Naturalist 170, 758–770.
| Changes in sperm quality and numbers in response to experimental manipulation of male social status and female attractiveness.Crossref | GoogleScholarGoogle Scholar | 17926297PubMed |
Della Togna, G, Trudeau, VL, Gratwicke, B, Evans, M, Augustine, L, Chia, H, Bronikowski, EJ, Murphy, JB, and Comizzoli, P (2017). Effects of hormonal stimulation on the concentration and quality of excreted spermatozoa in the critically endangered Panamanian golden frog (Atelopus zeteki). Theriogenology 91, 27–35.
| Effects of hormonal stimulation on the concentration and quality of excreted spermatozoa in the critically endangered Panamanian golden frog (Atelopus zeteki).Crossref | GoogleScholarGoogle Scholar | 28215683PubMed |
Dziminski, MA, Roberts, JD, and Simmons, LW (2010). Sperm morphology, motility and fertilisation capacity in the myobatrachid frog Crinia georgiana. Reproduction, Fertility and Development 22, 516–522.
| Sperm morphology, motility and fertilisation capacity in the myobatrachid frog Crinia georgiana.Crossref | GoogleScholarGoogle Scholar |
Guo, R, Zhang, W, Yang, Y, Ding, J, Ai, S, Yang, W, Bai, X, and Zhang, Y (2018). Sperm morphology and motility of Bufo raddei under long-term environmental heavy metal stress. Bulletin of Environmental Contamination and Toxicology 101, 305–313.
| Sperm morphology and motility of Bufo raddei under long-term environmental heavy metal stress.Crossref | GoogleScholarGoogle Scholar | 30069719PubMed |
Hettyey, A, Vági, B, Hévizi, G, and Török, J (2009). Changes in sperm stores, ejaculate size, fertilization success, and sexual motivation over repeated matings in the common toad, Bufo bufo (Anura: Bufonidae). Biological Journal of the Linnean Society 96, 361–371.
| Changes in sperm stores, ejaculate size, fertilization success, and sexual motivation over repeated matings in the common toad, Bufo bufo (Anura: Bufonidae).Crossref | GoogleScholarGoogle Scholar |
Hinkson, KM, and Poo, S (2020). Inbreeding depression in sperm quality in a critically endangered amphibian. Zoo Biology 39, 197–204.
| Inbreeding depression in sperm quality in a critically endangered amphibian.Crossref | GoogleScholarGoogle Scholar | 32141645PubMed |
Houlahan, JE, Findlay, CS, Schmidt, BR, Meyer, AH, and Kuzmin, SL (2000). Quantitative evidence for global amphibian population declines. Nature 404, 752–755.
| Quantitative evidence for global amphibian population declines.Crossref | GoogleScholarGoogle Scholar | 10783886PubMed |
Huo, L-J, Ma, X-H, and Yang, Z-M (2002). Assessment of sperm viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-term storage. Theriogenology 58, 1349–1360.
| Assessment of sperm viability, mitochondrial activity, capacitation and acrosome intactness in extended boar semen during long-term storage.Crossref | GoogleScholarGoogle Scholar | 12387348PubMed |
Kouba, AJ, Vance, CK, and Willis, EL (2009). Artificial fertilization for amphibian conservation: current knowledge and future considerations. Theriogenology 71, 214–227.
| Artificial fertilization for amphibian conservation: current knowledge and future considerations.Crossref | GoogleScholarGoogle Scholar | 19026442PubMed |
Kouba, AJ, Lloyd, RE, Houck, ML, Silla, AJ, Calatayud, N, Trudeau, VL, Clulow, J, Molinia, F, Langhorne, C, Vance, C, Arregui, L, Germano, J, Lermen, D, and Della Togna, G (2013). Emerging trends for biobanking amphibian genetic resources: The hope, reality and challenges for the next decade. Biological Conservation 164, 10–21.
| Emerging trends for biobanking amphibian genetic resources: The hope, reality and challenges for the next decade.Crossref | GoogleScholarGoogle Scholar |
Langhorne, CJ, Calatayud, NE, Kouba, CK, Willard, ST, Smith, T, Ryan, PL, and Kouba, AJ (2021). Efficacy of hormone stimulation on sperm production in an alpine amphibian (Anaxyrus boreas boreas) and the impact of short-term storage on sperm quality. Zoology 146, 125912.
| Efficacy of hormone stimulation on sperm production in an alpine amphibian (Anaxyrus boreas boreas) and the impact of short-term storage on sperm quality.Crossref | GoogleScholarGoogle Scholar | 33743452PubMed |
Larson, JL, and Miller, DJ (1999). Simple histochemical stain for acrosomes on sperm from several species. Molecular Reproduction and Development 52, 445–449.
| Simple histochemical stain for acrosomes on sperm from several species.Crossref | GoogleScholarGoogle Scholar | 10092125PubMed |
Lipke, C, Meinecke-Tillmann, S, and Meinecke, B (2009). Induced spermiation and sperm morphology in a dendrobatid frog, Dendrobates auratus (Amphibia, Anura, Dendrobatidae). Salamandra 45, 65–74.
Magnusson A, Skaug H, Neilsen A, Berg C, Kristensen K, Maechler M, van Bentham K, Bolker B, Sadat N, Ludecke D, Lenth R, O’Brien J, Brooks M (2020) ‘Generalized linear mixed models using template model builder: package ‘glmmTMB’.’ (CRAN)
Mansour, N, Lahnsteiner, F, and Patzner, RA (2009). Optimization of the cryopreservation of African clawed frog (Xenopus laevis) sperm. Theriogenology 72, 1221–1228.
| Optimization of the cryopreservation of African clawed frog (Xenopus laevis) sperm.Crossref | GoogleScholarGoogle Scholar | 19766299PubMed |
Maria, AN, Azevedo, HC, Santos, JP, Silva, CA, and Carneiro, PCF (2010). Semen characterization and sperm structure of the Amazon tambaqui Colossoma macropomum. Journal of Applied Ichthyology 26, 779–783.
| Semen characterization and sperm structure of the Amazon tambaqui Colossoma macropomum.Crossref | GoogleScholarGoogle Scholar |
Mayorga-Torres, JM, Agarwal, A, Roychoudhury, S, Cadavid, A, and Cardona-Maya, WD (2016). Can a short term of repeated ejaculations affect seminal parameters? Journal of Reproductive Infertility 17, 177–183.
McDonough, CE, Martin, MW, Vance, CK, Cole, JA, and Kouba, AJ (2016). Frequency of exogenous hormone therapy impacts spermiation in male Fowler’s toad (Bufo fowleri). Reproduction, Fertility and Development 28, 995–1003.
| Frequency of exogenous hormone therapy impacts spermiation in male Fowler’s toad (Bufo fowleri).Crossref | GoogleScholarGoogle Scholar |
Obringer, AR, O’Brien, JK, Saunders, RL, Yamamoto, K, Kikuyama, S, and Roth, TL (2000). Characterization of the spermiation response, luteinizing hormone release and sperm quality in the American toad (Bufo americanus) and the endangered Wyoming toad (Bufo baxteri). Reproduction, Fertility and Development 12, 51–58.
| Characterization of the spermiation response, luteinizing hormone release and sperm quality in the American toad (Bufo americanus) and the endangered Wyoming toad (Bufo baxteri).Crossref | GoogleScholarGoogle Scholar |
R Core Team (2017) ‘R: A language and environmental for statistical computing.’ (CRAN)
Roth TL, Obringer AR (2003) Reproductive research and the worldwide amphibian extinction crisis. In ‘Reproductive science and integrated conservation’. (Eds W Holt, A Pickard, J Rodger, D Wildt) pp. 359–374. (The Press Syndicate of the University of Cambridge: Cambridge, UK)
RStudio Team (2016) ‘RStudio: integrated development for R.’ (CRAN). Available at www.rstudio.com
Rubolini, D, Galeotti, P, Pupin, F, Sacchi, R, Nardi, PA, and Fasola, M (2007). Repeated matings and sperm depletion in the freshwater crayfish Austropotamobius italicus. Freshwater Biology 52, 1898–1906.
| Repeated matings and sperm depletion in the freshwater crayfish Austropotamobius italicus.Crossref | GoogleScholarGoogle Scholar |
Scarano, WR, Messias, AG, Oliva, SU, Klinefelter, GR, and Kempinas, WG (2006). Sexual behaviour, sperm quantity and quality after short-term streptozotocin-induced hyperglycaemia in rats. International Journal of Andrology 29, 482–488.
| Sexual behaviour, sperm quantity and quality after short-term streptozotocin-induced hyperglycaemia in rats.Crossref | GoogleScholarGoogle Scholar | 16524366PubMed |
Silla, AJ, and Byrne, PG (2019). The role of reproductive technologies in amphibian conservation breeding programs. Annual Review of Animal Biosciences 7, 499–519.
| The role of reproductive technologies in amphibian conservation breeding programs.Crossref | GoogleScholarGoogle Scholar | 30359086PubMed |
Silla, AJ, and Roberts, JD (2012). Investigating patterns in the spermiation response of eight Australian frogs administered human chorionic gonadotropin (hCG) and luteinizing hormone-releasing hormone (LHRHa). General and Comparative Endocrinology 179, 128–136.
| Investigating patterns in the spermiation response of eight Australian frogs administered human chorionic gonadotropin (hCG) and luteinizing hormone-releasing hormone (LHRHa).Crossref | GoogleScholarGoogle Scholar | 22909973PubMed |
Silla, AJ, McFadden, MS, and Byrne, PG (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, coy080.
| Hormone-induced sperm-release in the critically endangered Booroolong frog (Litoria booroolongensis): effects of gonadotropin-releasing hormone and human chorionic gonadotropin.Crossref | GoogleScholarGoogle Scholar | 30792859PubMed |
Stuart, SN, Chanson, JS, Cox, NA, Young, BE, Rodrigues, ASL, Fischman, DL, and Waller, RW (2004). Status and trends of amphibian declines and extinctions worldwide. Science 306, 1783–1786.
| Status and trends of amphibian declines and extinctions worldwide.Crossref | GoogleScholarGoogle Scholar | 15486254PubMed |
Swerdloff, RS, Handelsman, DJ, and Bhasin, S (1985). Hormonal effects of GnRH agonist in the human male: an approach to male contraception using combined androgen and GnRH agonist treatment. Journal of Steroid Biochemistry 23, 855–861.
| Hormonal effects of GnRH agonist in the human male: an approach to male contraception using combined androgen and GnRH agonist treatment.Crossref | GoogleScholarGoogle Scholar | 2934582PubMed |
Trudeau, VL, Somoza, GM, Natale, GS, Pauli, B, Wignall, J, Jackman, P, Doe, K, and Schueler, FW (2010). Hormonal induction of spawning in 4 species of frogs by coinjection with a gonadotropin-releasing hormone agonist and a dopamine antagonist. Reproductive Biology and Endocrinology 8, 36.
| Hormonal induction of spawning in 4 species of frogs by coinjection with a gonadotropin-releasing hormone agonist and a dopamine antagonist.Crossref | GoogleScholarGoogle Scholar | 20398399PubMed |
Uteshev, VK, Shishova, NV, Kaurova, SA, Browne, RK, and Gakhova, EN (2012). Hormonal induction of spermatozoa from amphibians with Rana temporaria and Bufo bufo as anuran models. Reproduction, Fertility and Development 24, 599–607.
| Hormonal induction of spermatozoa from amphibians with Rana temporaria and Bufo bufo as anuran models.Crossref | GoogleScholarGoogle Scholar |
Villaverde-Morcillo, S, Soler, AJ, Esteso, MC, Castaño, C, Miñano-Berna, A, Gonzalez, F, and Santiago-Moreno, J (2017). Immature and mature sperm morphometry in fresh and frozen-thawed falcon ejaculates. Theriogenology 98, 94–100.
| Immature and mature sperm morphometry in fresh and frozen-thawed falcon ejaculates.Crossref | GoogleScholarGoogle Scholar | 28601162PubMed |
Wlizla, M, Falco, R, Peshkin, L, Parlow, AF, and Horb, ME (2017). Luteinizing hormone is an effective replacement for hCG to induce ovulation in Xenopus. Developmental Biology 426, 442–448.
| Luteinizing hormone is an effective replacement for hCG to induce ovulation in Xenopus.Crossref | GoogleScholarGoogle Scholar | 27263125PubMed |
Wolf, DP, and Hedrick, JL (1971). A molecular approach to fertilization: II. Viability and artificial fertilization of Xenopus laevis gametes. Developmental Biology 25, 348–359.
| A molecular approach to fertilization: II. Viability and artificial fertilization of Xenopus laevis gametes.Crossref | GoogleScholarGoogle Scholar | 5567826PubMed |
Zohar, Y, and Mylonas, CC (2001). Endocrine manipulations of spawning in cultured fish: from hormones to genes. Aquaculture 197, 99–136.
| Endocrine manipulations of spawning in cultured fish: from hormones to genes.Crossref | GoogleScholarGoogle Scholar |
