Biological characterization of rodent and human vasopressin V1breceptors using SSR-149415, a nonpeptide V1breceptor ligand
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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- 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)
- ORCID
- Allen, John
- Venue
- American Journal of Physiology-Regulatory, Integrative and Comparative Physiology
- Publisher
- American Physiological Society
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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