Pseudogenization of Mc1r gene associated with transcriptional changes related to melanogenesis explains leucistic phenotypes in Oreonectes cavefish (Cypriniformes, Nemacheilidae)

Abstract

Organisms that have colonized underground caves encounter vastly different selective pressures than their relatives in above‐ground habitats. While disruption of certain pigmentation genes has been documented in various cave‐dwelling taxa, little is known about wider impacts across pigmentation and other gene pathways. We here study the timeframe and transcriptional landscape of a leucistic and blind cypriniform fish (Oreonectes daqikongensis, Nemacheilidae) that inhabits karst caves in Guizhou, China. Based on data from the mitochondrial ND4, ND5, and Cytb genes, we show that the divergence between O. daqikongensis and its most closely related pigmented species occurred ca. 6.82 million years ago (95% HPD, 5.12–9.01), providing ample time for widespread phenotypic change. Indeed, we found that the DNA sequence of Mc1r (melanocortin‐1 receptor), a key gene regulating the biosynthesis of melanin in most vertebrates, is pseudogenized in O. daqikongensis, caused by a 29 bp deletion in the protein‐coding region. Furthermore, 99,305 unigenes were annotated based on the transcriptome of skin tissue of Oreonectes fish. Among the differentially expressed unigenes, 7,326 (7.4% of the total unigenes) had decreased expression and 2,530 (2.5% of the total unigenes) had increased expression in O. daqikongensis skin. As predicted, the expression of Mc1r and 18 additional genes associated with melanin biosynthesis was significantly downregulated in the skin tissue of O. daqikongensis, but not in its congener. Our results, integrating with other studies on cavefishes, suggest that loss of pigmentation was caused by coding region loss‐of‐function mutations along with widespread transcriptional changes, resulting from extended evolutionary time as a cave‐dwelling form.