Heterochronic development of pelvic fins in zebrafish: possible involvement of temporal regulation of pitx1 expression.

Anterior and posterior paired appendages of vertebrates are notable examples of heterochrony in the relative timing of their development. In teleosts, posterior paired appendages (pelvic fin buds) emerge much later than their anterior paired appendages (pectoral fin buds). Pelvic fin buds of zebrafish (Danio rerio) appear at 3 weeks post-fertilization (wpf) during the larva-to-juvenile transition (metamorphosis), whereas pectoral fin buds arise from the lateral plate mesoderm on the yolk surface at the embryonic stage. Here we explored the mechanism by which presumptive pelvic fin cells maintain their fate, which is determined at the embryonic stage, until the onset of metamorphosis. Expression analysis revealed that transcripts of pitx1, one of the key factors for the development of posterior paired appendages, became briefly detectable in the posterior lateral plate mesoderm at early embryonic stages. Further analysis indicated that the pelvic fin-specific pitx1 enhancer was in the poised state at the larval stage and is activated at the juvenile stage. We discuss the implications of these findings for the heterochronic development of pelvic fin buds.

Euryhaline fish larvae ingest more microplastic particles in seawater than in freshwater.

Microplastic (MP) pollution is a major concern in aquatic environments. Many studies have detected MPs in fishes; however, little is known about differences of microplastic uptake by fish in freshwater (FW) and those in seawater (SW), although physiological conditions of fish differ significantly in the two media. In this study, we exposed larvae (21 days post-hatching) of Oryzias javanicus (euryhaline SW) and Oryzias latipes (euryhaline FW), to 1-µm polystyrene microspheres in SW and FW for 1, 3, or 7 days, after which, microscopic observation was conducted. MPs were detected in the gastrointestinal tracts in both FW and SW groups, and MP numbers were higher in the SW group in both species. Vertical distribution of MPs in the water, and body sizes of both species exhibited no significant difference between SW and FW. Detection of water containing a fluorescent dye revealed that O. javanicus larvae swallowed more water in SW than in FW, as has also been reported for O. latipes. Therefore, MPs are thought to be ingested with water for osmoregulation. These results imply that SW fish ingest more MPs than FW fish when exposed to the same concentration of MPs.

Localization of ß-Catenin and Islet in the Pelvic Fin Field in Zebrafish.

In zebrafish, pelvic fin buds appear at 3 weeks post fertilization (wpf) during the larval to juvenile transition (metamorphosis), but their fate is already determined during embryogenesis. Thus, presumptive pelvic fin cells appear to memorize their positional information for three weeks, but no factors expressed in the pelvic fin field from the embryonic to the metamorphic stages have been identified. In mice, Islet1 is proposed to promote nuclear accumulation of ß-catenin in the hindlimb field, which leads to the initiation of hindlimb bud outgrowth through activation of the Wnt/ßcatenin pathway. Here, we examined the distribution of ß-catenin and islet proteins in the pelvic fin field of zebrafish from the embryonic to the metamorphic stages. We found that transcripts of islet2a, but not islet1, are detected in the posterior lateral plate mesoderm, including the presumptive pelvic fin field, at the embryonic stage as well as in the pelvic fin bud at the metamorphic stage. Immunolocalization revealed that ß-catenin and islet proteins, which are synthesized during the embryonic stage, remain in the cytoplasm of the presumptive pelvic fin cells during the larval stage, and are then translocated into the nuclei of the pelvic fin bud at the metamorphic stage. We propose that cytoplasmic localization of these proteins in the presumptive pelvic fin cells that remained during the larval stage may underlie the mechanism by which pelvic fin cells memorize their positional information from the embryonic stage to the metamorphic stage.