Expressão gênica durante o desenvolvimento ocular e regulação de assimetria de opsinas na espécie Anableps anableps, peixe de quatro olhos

Abstract

The ocular development is a complex process orchestrated by several events that include: cell specification, morphogenesis and differentiation. All these developmental and functional processes are extremely conserved among living vertebrate species, however, unique adaptations are sometimes observed, such as in fish of the genus Anableps. Unlike most fishes, which use their eyes to explore a submerged world, in Anableps anableps (Anablepidae: Cyprinodontiformes), the eye is adapted for the simultaneous perception of a world above and below the water line. These exceptional adaptations include: duplicated corneas and pupils, as well as a specialized retina region associated with simultaneous aerial and aquatic vision, and which have asymmetrical gene expression. Recently, by transcriptomic analysis of the developing eyes of A. anableps, 20 non-visual opsin genes were identified to be asymmetrically expressed between pre- and post-duplication stages of corneas and pupils. Thus, here, we analyzed by in situ hybridization the gene expression of a bistable opsin (parapinopsin) and a neuropsin (Opn5) in A. anableps larvae. Our data showed that the pattern of gene expression of these opsins is symmetrical between the dorsal and ventral retina, respectively, with expression in the ONL, INL and GCL layers. We also investigated the expression of three non-visual melanopsin genes (opn4x1, opn4x2, opn4m3), one eleost multiple tissue opsin (tmt1b), and two visual opsins (lws and rh2-1) in the dorsal and ventral retinas of juvenile A. anableps, after changing the photic conditions in which the juvenile fish were. Then, we showed that in the transition from an environment of high turbidity to one of clear water, opsins have their expression patterns changed. Additionally, by immunofluorescence, we revealed the expression of Lamin A/C, proteins expressed as part of eye development in A. anableps as well as in other developing organisms, as well as in adult individuals. Therefore, we believe that the information described here elucidates many aspects of the molecular mechanisms behind the development and adaptive plasticity of A. anableps’ eyes.