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. 2019 Dec 17;9:19295. doi: 10.1038/s41598-019-55675-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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I8-arachnotocin is a competitive antagonist at V_1aR. (a) Accumulation of second messengers (IP₁ and cAMP) with partial activating concentrations of endogenous ligands (50 nM OT at OTR; 1 nM VP at V_1aR; 3 nM VP at V_1bR; 0.5 pM at V₂R) in the absence and presence of 1 or 10 µM I8-arachnotocin in comparison to a saturating concentration of endogenous ligand (10 µM). All data were normalized to second messenger accumulation above baseline (0%) and maximum (100%) activity of the endogenous ligand. The dashed line depicts IP₁/cAMP accumulation in the absence of I8-arachnotocin. Error bars depict SEM. n = 3, except n = 2 for V_1aR (error bars depict SD). The asterisk (*) indicates significance in Student’s t-test (p = 0.0226). (b) Accumulation of IP₁ by stimulation of human V_1aR with VP (10 pM – 10 µM) alone or in the presence of 1, 3 or 10 µM I8-arachnotocin. Receptor activation was normalized to the accumulation of IP₁ above baseline. Data points were normalized to the maximum (100%) and minimum (0%) response generated by VP. Error bars depict SEM; n = 3. Insert: Schild regression analysis: A = EC₅₀ of VP in presence of I8-arachnotocin; A′ = EC₅₀ of VP; B = logarithm of I8-arachnotocin concentration; Schild slope 1.28 ± 0.02 (SEM), R² = 0.9998. The dotted line represents a reference line with a slope of 1; n = 3.