Cited by 35Neuropeptides and Animal PhysiologyNeuroendocrine regulation and behaviorReceptor Mechanisms and Signaling

Biological characterization of rodent and human vasopressin V1breceptors using SSR-149415, a nonpeptide V1breceptor ligand

Gal, Claudine Serradeil-Le, Raufaste, Danièle, Derick, Sylvain, Blankenstein, Jörg, Allen, John, Pouzet, Brigitte, Pascal, Marc, Wagnon, Jean +1 more·2007·American Journal of Physiology-Regulatory, Integrative and Comparative Physiology

Abstract

[(3)H]SSR-149415 is the first tritiated nonpeptide vasopressin V(1b) receptor (V(1b)R) antagonist ligand. It was used for studying rodent (mouse, rat, hamster) and human V(1b)R from native or recombinant origin. Moreover, a close comparison between the human and the mouse V(1b)R was performed using SSR-149415/[(3)H]SSR-149415 in binding and functional studies in vitro. [(3)H]SSR-149415 binding was time-dependent, reversible, and saturable. Scatchard plot analysis gave a single class of high-affinity binding sites with apparent equilibrium dissociation constant (K(d)) approximately 1 nM and maximum binding density (B(max)) values from 7,000 to 300,000 sites/cell according to the cell line. In competition experiments, [(3)H]SSR-149415 binding was stereospecific and dose-dependently displaced by reference peptide and nonpeptide arginine vasopressin (AVP)/OT ligands following a V(1b) rank order of affinity: SSR-149415 = AVP > dCha > dPen > dPal > dDavp > SSR-126768A > SR-49059 > SSR-149424 > OT > SR-121463B. Species differences between human, rat, mouse, and hamster V(1b)R were observed. Autoradiography studies with [(3)H]SSR-149415 on rat and human pituitary showed intense specific labeling confined to corticotroph cells and absence of labeling in the other tissues examined. SSR-149415 potently and stereospecifically antagonized the AVP-induced inositol phosphate production and intracellular Ca(2+) increase (EC(50) from 1.83 to 3.05 nM) in recombinant cell lines expressing either the mouse or the human V(1b)R. AVP (10(-7) M) exposure of AtT20 cells expressing mouse or human EGFP-tagged V(1b)R induced their rapid internalization. Preincubation with 10(-6) M SSR-149415 counteracted the internalization process. Moreover, recycling of internalized receptors was observed upon 10(-6) M SSR-149415 treatment. Thus SSR-149415/[(3)H]SSR-149415 are unique tools for studying animal and human V(1b)R.

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Crossref · 10.1152/ajpregu.00062.2007

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Authors
Gal, C. S.-L., Raufaste, D., Derick, S., Blankenstein, J., Allen, J., Pouzet, B., Pascal, M., Wagnon, J., & Ventura, M. A.
Affiliations
Sanofi (France) (FR)
Venue
American Journal of Physiology-Regulatory, Integrative and Comparative Physiology
Published
2007-08
Volume
293
Issue
2
Pages
R938-R949
ISSN
  • 0363-6119 (ISSN-L)
  • 1522-1490
OpenAlex
W1992214677
Cited by
35
References
51
Citations by year
2026: 2 · 2024: 2 · 2023: 1 · 2022: 2 · 2021: 2 · 2020: 1 · 2017: 4 · 2016: 2 · 2015: 1 · 2014: 2 · 2013: 3 · 2012: 2
FWCI
1.05
Type
journal article
Language
en
Open access
closed
Locations
  • American Journal of Physiology-Regulatory, Integrative and Comparative Physiology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Institut National de la Santé et de la Recherche Médicale
  • Centre National de la Recherche Scientifique
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
  • Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
receptor internalization, vasopressin V1b receptor, non-peptide antagonists, vasopressin, oxytocin, receptor desensitization, receptor resensitization, corticotrophs, inositol phosphate formation, intracellular calcium
MeSH terms
Animals, Autoradiography, Calcium, Cell Line, Endocytosis, Cricetinae, Humans, Indoles, Inositol Phosphates, Ligands, Membrane Proteins, Pyrrolidines, Radioligand Assay, Recombinant Proteins, Sensitivity and Specificity, Tritium, Receptors, Vasopressin, DNA, Complementary, Mice

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