Urotensin II-related peptide (Urp) is expressed in motoneurons in zebrafish, but is dispensable for locomotion in larva

Urp1 and Urp2 act redundantly to maintain spine shape in zebrafish larvae

“…Thus, to test the idea that Urp1/2 signaling acts on muscle, we aimed at blocking muscle contractions with myosin II inhibitors (BTS and blebbistatin) and assayed the consequences on Urp2-induced tail bending in embryos. We have previously shown that a heat-shock-driven overexpression of urp2 in embryos at 1 dpf induces an upward bending of the tail (Quan et al, 2021). Strikingly, when embryos were treated with blebbistatin prior to the heat shock, the Urp2-induced bending was abolished (Fig.…”

“…Nevertheless, several other studies suggest that Urp1/2-Uts2r3 signaling is functional during embryogenesis. On the one hand, overexpressing Urp1 or Urp2 by different means in 1 dpf embryos results in a backward bending of the tail (Cantaut-Belarif et al, 2020; Lu et al, 2020; Quan et al, 2021; Zhang et al, 2018). On the other hand, the expression of urp1 and/or urp2 is reduced in zmind and sspo mutants and injection of urp1 mRNA can rescue their phenotypes (Cantaut-Belarif et al, 2020; Lu et al, 2020; Zhang et al, 2018).…”

“…The hsp:urp2 tol2 transgenesis vector was already described (Quan et al, 2021). The ubi:urp2 and ubi:STOP lox -urp2 tol2 transgenesis vectors were produced using plasmids from the tol2kit (Kwan et al, 2007) and Gateway recombination system (Life Technologies).…”

“…To produce the entry plasmid p5E-ubi:STOP lox ( pCM351 ), the loxP -flanked STOP cassette from pDH083 (Hesselson et al, 2009) was PCR-amplified with primers starting at the 5’ end of the loxP sequences with added BglII restriction sites, cut with BglII, ligated into BamHI-linearized pENTR5’_ubi ( pCM206 ) (Mosimann et al, 2011), and the resulting clones verified by restriction digest and sequencing. Then, p5Eubi:urp2 (Ref) or p5E -ubi:STOP lox , pME- urp2 (Quan et al, 2021) and p3E-polyA were recombined into the pDestTol2CG2 destination vector. For transgenesis, solutions containing transgenesis plasmids (30ng/µl) and tol2 transposase mRNA (25 ng/µl) were prepared and injected (2 nl) into 1 or 2 cell-stage embryos.…”

“…Characterized first in the Japanese eel (Anguilla) (Nobata et al, 2011) and in the zebrafish (Parmentier et al, 2011), urp1 and urp2 seem to exist in all fishes but to have been lost in mammals. However, urp2 has recently also been identified in Xenopus (clawed frogs) (Alejevski et al, 2021). All peptides of the Uts2 family have been suggested to act through a multigenic family of G protein-coupled receptors called Uts2r (also known as Utr or UT).…”