Sexual signalling using dynamic skin colouration is a key feature in some vertebrates; however, it is rarely studied in amphibians. Consequently, little is known about the hormonal basis of this interesting biological phenomenon for many species. Male stony creek frogs (Litoria wilcoxii) are known to change dorsal colouration from brown to lemon yellow within minutes. This striking change is faster then what has been seen most amphibians, and could therefore be under neuronal regulation, a factor that is rarely observed in amphibians. In this study, we observed colour changes in wild frogs during amplexus to determine the natural timing of colour change. We also investigated the hypothesis that colour change is mediated by either reproductive or neuro- hormones. This was achieved by injecting frogs with epinephrine, testosterone, saline solution (control 1) or sesame oil (control 2). A non-invasive approach was also used wherein hormones and controls were administered topically. Male frogs turned a vivid yellow within 5 minutes of initiation of amplexus and remained so for 3–5 hours before rapidly fading back to brown. Epinephrine-treated frogs showed a significant colour change from brown to yellow within 5 minutes, however, testosterone-treated frogs did not change colour. Our results provide evidence of the role neuronal regulation plays in colour change systems.
Rapid colour changes in vertebrates have fascinated biologists for centuries, herein we demonstrate dynamic colour change in an anuran amphibian, the stony creek frog (Litoria wilcoxii), which turns from brown to bright (lemon) yellow during amplexus. We show this by comparing the colour of baseline (unpaired males) and amplecting (paired) males. We also investigate the possible role of stress and stress hormones on this colour change. Frogs were subjected to four different levels of stressors (handling, toe-clipping, saline injection and adrenocorticotropic hormone [ACTH] injection) and the colour change was measured using digital photography. A comparison of baseline colour and stress hormone (corticosterone) levels was also conducted to give further insight to this topic. From the images, the Red Blue Green (RGB) colour values were calculated, and a principal components analysis (PCA) was used to create a single colour metric (the major axis) as an index of colour in the visible spectrum. A moderate stressor (toe-clipping) led to a significant change in colour (within 10 min) similar to that of amplecting males. Surprisingly, neither a mild stressor (handling and saline injection) nor the maximum stressor (handling and ACTH injection) led to a lightening response. This study confirms that the dynamic male colour change in this species in response to medium stressors adds new knowledge to the understanding of the functional mechanisms of dynamic colour change in amphibians.
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