New sources of retinoic acid synthesis revealed by live imaging of an Aldh1a2-GFP reporter fusion protein throughout zebrafish development.

BACKGROUND: Regulation of synthesis and turnover of retinoic acid (RA) is an important mechanism that controls the activity of RA signaling during vertebrate development. During embryonic patterning, the dynamic expression patterns of the aldh1a2 gene, which encodes a retinaldehyde dehydrogenase, provide the major source of RA, whereas the only other retinaldehyde dehydrogenase in teleosts, aldh1a3, is expressed later and locally restricted. Aldh1a2-mediated RA synthesis has been shown to also regulate adult cell fates, such as during heart and fin regeneration. However, only very few other sites of postembryonic RA synthesis in vertebrates are known. We generated transgenic lines in zebrafish by BAC recombineering that express a fusion protein of Aldh1a2 and green fluorescent protein (GFP) under the control of endogenous aldh1a2 regulatory sequences (aldh1a2:gfp). RESULTS: aldh1a2:gfp reports the complete endogenous expression pattern in embryos and rescues embryonic lethality in aldh1a2 mutants. We identify novel postembryonic sources of RA synthesis, including lateral line support cells, in kidney-derived organs that regulate calcium homeostasis, and in perichordal cells during vertebral development. CONCLUSIONS: The novel aldh1a2 reporter line is driven by the complete set of regulatory sequences required for zebrafish development, reports novel sources of RA synthesis, and identifies the source of RA that promotes vertebral ossification.

An organizer controls the development of the "sword," a sexually selected trait in swordtail fish.

Male swordtail fish of the genus Xiphophorus (Poeciliidae) possess a "sword" that is composed of several colored elongated ventral fin rays of the caudal fin. The sword is a secondary sexual trait that evolved through sexual selection by female preference. To uncover the developmental mechanisms underlying the metamorphosis from a juvenile caudal fin to the sword, we have devised a transplantation protocol to assay the fate of single transplanted fin rays and their interactions with flanking rays. These experiments provide evidence for the existence of a previously unrecognized inductive signal that originates in those rays that develop into the two longest sword rays. This "sword organizer" causes adjacent fin rays to grow and become integrated into the sword and induces the development of an additional, typically pigmented sword in grafts to the dorsal part of the caudal fin. We show that the potential to develop a sword is restricted to certain parts of the caudal fin. Our findings suggest that the evolution of swords in swordtails required the acquisition of two developmental mechanisms: the establishment of signaling competence in prospective sword rays in the embryo or early larva, and its activation through androgen signaling in adult male fish.

Expression of zebrafish aldh1a3 (raldh3) and absence of aldh1a1 in teleosts.

The vitamin A-derived morphogen retinoic acid (RA) plays important roles during the development of chordate animals. The Aldh1a-family of RA-synthesizing enzymes consists of three members, Aldh1a1-3 (Raldh1-3), that are dynamically expressed throughout development. We have searched the known teleost genomes for the presence of Raldh family members and have found that teleost fish possess orthologs of Aldh1a2 and Aldh1a3 only. Here we describe the expression of aldh1a3 in the zebrafish, Danio rerio. Whole mount in situ hybridization shows that aldh1a3 is expressed during eye development in the retina flanking the optic stalks and later is expressed ventrally, opposite the expression domain of aldh1a2. During inner ear morphogenesis, aldh1a3 is expressed in developing sensory epithelia of the cristae and utricular macula and is specifically up-regulated in epithelial projections throughout the formation of the walls of the semicircular canals and endolymphatic duct. In contrast to the mouse inner ear, which expresses all three Raldhs, we find that only aldh1a3 is expressed in the zebrafish otocyst, while aldh1a2 is present in the periotic mesenchyme. During larval stages, additional expression domains of aldh1a3 appear in the anterior pituitary and the swim bladder. Our analyses provide a starting point for genetic studies to examine the role of RA in these organs and emphasize the suitability of the zebrafish inner ear in dissecting the contribution of RA signaling to the development of the vestibular system.