Abstract

BackgroundThe non-lethal collection of sperm from live males is an important component for multiple captive-breeding techniques, including assisted reproductive technology (ART) protocols, sperm cryopreservation and in vitro fertilization. However, in amphibians, the type and amount of hormone necessary to induce spermiation can be highly variable, even among closely related species. We are unaware of any studies that have examined the spermiation response to exogenous hormones across highly differentiated populations within a species.MethodsWe examined variation in sperm viability and production in response to the hormone LHRH among four divergent populations of the Red-eyed Treefrog (Agalychnis callidryas). We hypothesized that these highly differentiated populations would show variability in sperm count and viability in response to two dosages, 2 μg/g and 4 μg/g, of the hormone LHRH. We collected spermic urine 3 h post injection (PI). We then examined variation in spermiation at 3, 7, 12, and 24 h PI of LHRH for two allopatric populations that previously showed evidence of premating behavioral isolation.ResultsOne population of Red-eyed Treefrog exhibited reduced sperm viability but not count in response to the hormone LHRH compared to all other populations. In addition, we found peak viability at 3 h PI for the allopatric population comparison. There was no difference in sperm production within or between populations at 3, 7, 12, or 24 h PI. For both studies, intrapopulation variation was high.ConclusionART often focuses on threatened species with small, isolated populations, which could evolve localized differences due to the evolutionary process of drift and selection. The high variation in response and the population-level differences in sperm viability we observed demonstrate that practitioners of ART should consider the possibility of divergent responses to hormones which may affect study design and animal receptivity to ART protocols.Electronic supplementary materialThe online version of this article (doi:10.1186/s12958-016-0216-3) contains supplementary material, which is available to authorized users.

Highlights

  • The non-lethal collection of sperm from live males is an important component for multiple captivebreeding techniques, including assisted reproductive technology (ART) protocols, sperm cryopreservation and in vitro fertilization

  • The results did not change with body weight dropped from the model: we detected no difference in sperm count in response to either dose of Luteinizing hormone-release hormone (LHRH) among or within (F3,45 = 0.55, P = 0.46) populations

  • We found that exogenous hormone LHRH was effective in producing viable sperm from all populations of Red-eyed Treefrogs in our study

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Summary

Introduction

The non-lethal collection of sperm from live males is an important component for multiple captivebreeding techniques, including assisted reproductive technology (ART) protocols, sperm cryopreservation and in vitro fertilization. We are unaware of any studies that have examined the spermiation response to exogenous hormones across highly differentiated populations within a species. Many ART protocols for amphibians combine traditional breeding with laboratory methods (e.g., the use of hormones) for successful breeding and induction of viable oocytes and sperm for cryopreservation and/or in vitro fertilization [4, 5]. ART protocols used sperm that were collected from testes macerates, sacrificing males [6,7,8], within the past 20 years, exogenous hormones have been used to obtain sperm non-invasively [9]. At least 12 species of anurans representing seven families have been bred using ART [10]

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