Identifying and locating carotenoids in supra-orbital combs of male black grouse (Lyurus tetrix) using Raman and transmission electron microscopy: A histological study using rehydrated tissue samples

Colourful signals have long been implicated as indicators of individual quality in animals. Bare-skin signals are an understudied aspect of avian colouration compared with plumage studies, despite displaying rapid changes in size and colour in response to different environmental or physiological stressors. Even fewer studies have focused on the underlying histology of these structures and the importance this plays in the resulting skin colour. Using the Black Grouse (Lyrurus tetrix), we identified the underlying structure of individual dermal spikes, which make up the red supra-orbital comb (a known integumentary signal of male quality), and highlight visual structural differences between combs of different sizes. In addition, we used Raman spectroscopy to indicate the presence of carotenoids within the tissue, something that had previously only been inferred through characteristic reflectance patterns. An increased understanding of the structural basis of colour of featherless parts of the skin opens up exciting new avenues for interpreting the information content of integumentary signals.