Comparative distribution and in vitro activities of the urotensin II-related peptides URP1 and URP2 in zebrafish: evidence for their colocalization in spinal cerebrospinal fluid-contacting neurons

Feng B Quan; Christophe Dubessy; Sonya Galant; Natalia B Kenigfest; Lydia Djenoune; Jérôme Leprince; Claire Wyart; Isabelle Lihrmann; Hervé Tostivint

doi:10.1371/journal.pone.0119290

Comparative distribution and in vitro activities of the urotensin II-related peptides URP1 and URP2 in zebrafish: evidence for their colocalization in spinal cerebrospinal fluid-contacting neurons

PLoS One. 2015 Mar 17;10(3):e0119290. doi: 10.1371/journal.pone.0119290. eCollection 2015.

Authors

Feng B Quan¹, Christophe Dubessy², Sonya Galant³, Natalia B Kenigfest⁴, Lydia Djenoune⁵, Jérôme Leprince², Claire Wyart⁶, Isabelle Lihrmann², Hervé Tostivint¹

Affiliations

¹ Evolution des Régulations Endocriniennes, UMR 7221 CNRS, and Muséum National d'Histoire Naturelle, Paris, France.
² Inserm, U982, University of Rouen, Mont-Saint-Aignan, France; Laboratory of Neuronal and Neuroendocrine Differentiation and Communication, Institute for Research and Innovation in Biomedicine (IRIB), University of Rouen, Mont-Saint-Aignan, France; Normandy University, University of Rouen, Mont-Saint-Aignan, France.
³ Laboratoire de Neurobiologie et Développement, CNRS UPR 3294, Institut Alfred Fessard, Gif-sur-Yvette, France.
⁴ Evolution des Régulations Endocriniennes, UMR 7221 CNRS, and Muséum National d'Histoire Naturelle, Paris, France; Laboratory of Evolution of Neuronal Interactions, Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, Russia.
⁵ Evolution des Régulations Endocriniennes, UMR 7221 CNRS, and Muséum National d'Histoire Naturelle, Paris, France; Institut du Cerveau et de la Moelle épinière, ICM, Inserm U 1127, CNRS, UMR 7225, Sorbonne Universités, UPMC University Paris 06 UMR S 1127, Paris, France.
⁶ Institut du Cerveau et de la Moelle épinière, ICM, Inserm U 1127, CNRS, UMR 7225, Sorbonne Universités, UPMC University Paris 06 UMR S 1127, Paris, France.

Abstract

Urotensin II (UII) is an evolutionarily conserved neuropeptide initially isolated from teleost fish on the basis of its smooth muscle-contracting activity. Subsequent studies have demonstrated the occurrence of several UII-related peptides (URPs), such that the UII family is now known to include four paralogue genes called UII, URP, URP1 and URP2. These genes probably arose through the two rounds of whole genome duplication that occurred during early vertebrate evolution. URP has been identified both in tetrapods and teleosts. In contrast, URP1 and URP2 have only been observed in ray-finned and cartilaginous fishes, suggesting that both genes were lost in the tetrapod lineage. In the present study, the distribution of urp1 mRNA compared to urp2 mRNA is reported in the central nervous system of zebrafish. In the spinal cord, urp1 and urp2 mRNAs were mainly colocalized in the same cells. These cells were also shown to be GABAergic and express the gene encoding the polycystic kidney disease 2-like 1 (pkd2l1) channel, indicating that they likely correspond to cerebrospinal fluid-contacting neurons. In the hindbrain, urp1-expressing cells were found in the intermediate reticular formation and the glossopharyngeal-vagal motor nerve nuclei. We also showed that synthetic URP1 and URP2 were able to induce intracellular calcium mobilization in human UII receptor (hUT)-transfected CHO cells with similar potencies (pEC50=7.99 and 7.52, respectively) albeit at slightly lower potencies than human UII and mammalian URP (pEC50=9.44 and 8.61, respectively). The functional redundancy of URP1 and URP2 as well as the colocalization of their mRNAs in the spinal cord suggest the robustness of this peptidic system and its physiological importance in zebrafish.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Animals
CHO Cells
Cells, Cultured
Cerebrospinal Fluid / metabolism*
Cricetinae
Cricetulus
Embryo, Nonmammalian / cytology
Embryo, Nonmammalian / metabolism
Fluorescent Antibody Technique
Humans
In Situ Hybridization
Intracellular Signaling Peptides and Proteins
Neurons / cytology
Neurons / metabolism*
Peptide Fragments / metabolism*
Peptide Hormones / metabolism
RNA, Messenger / genetics
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Rhombencephalon / cytology
Rhombencephalon / metabolism*
Spinal Cord / cytology
Spinal Cord / metabolism*
Urotensins / genetics
Urotensins / metabolism*
Zebrafish / growth & development
Zebrafish / metabolism*

Substances

Intracellular Signaling Peptides and Proteins
Peptide Fragments
Peptide Hormones
RNA, Messenger
UTS2B protein, human
Urotensins
urotensin II

Grants and funding

This research was supported by funding obtained from the Centre National de la Recherche Scientifique and the Muséum National d’Histoire Naturelle to HT, and from the Institut de la Santé et de la Recherche Médicale, the University of Rouen and the Conseil Régional de Haute-Normandie to CD and IL. CW received financial support from the network Ecole des Neurosciences de Paris (ENP), the Fondation Bettencourt Schueller (FBS), Mr Pierre Belle, the City of Paris Emergence program, the Atip/Avenir junior program from INSERM and CNRS, the Fyssen foundation, the International Reintegration Grant from Marie Curie Actions Framework Program 6, and the European Research Council (ERC) starter grant “OptoLoco”. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.