AbstractPigments such as melanin are most often associated with the surface of an organism, providing functions such as coloration and protection from UV radiation. However, the internal organs of some species also contain melanin. Internal melanin may also perform protective functions when cellular stress is experienced. We tested liver tissue of two tree lizard species that experienced introgression of their mitochondria to see whether melanin was present and whether it was at higher concentrations in the types of lizards thought to be under cellular stress. Previous work found that mitochondria from livers of hybrid tree lizards (those with introgressed mitochondrial DNA) had higher levels of ATP production at higher temperatures than both parental species and showed indications of dysfunction. Therefore, we predicted that if internal melanin functions to mediate the impact of damaging by-products of metabolism, such as excessive reactive oxygen species (ROS), melanin would be highest in livers of lizards with introgressed mitochondria. To test this, we used a melanin-specific stain on liver tissue sections and measured melanin concentration with spectrophotometry of heavy-membrane fractions from whole-liver homogenates of both parental species and their hybrids with introgressed mitochondria. Slides of liver sections treated with a melanin-specific stain revealed that hybrids contained melanin and had significantly higher levels of it than either parental species. Spectrophotometry gave the same result. This distribution of internal melanin matches the pattern expected if melanin functions as a compensatory mechanism to deal with higher ATP production and the subsequent high levels of ROS expected in these hybrid lizards. Future studies will examine this mechanistically.
Keywords: ATP; hybrid lizards; internal melanin; liver; melanin; mitochondria; reactive oxygen species (ROS).