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. 2019 Apr 25;9:6523. doi: 10.1038/s41598-019-43002-2

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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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Cilioplasmic and cytoplasmic cAMP levels are differentially regulated. Peak cAMP increase in response to each stimulus is summarized in the bar graph for (a) cilioplasm and (b) cytoplasm. N = 4 experiments for each group. *P < 0.05, **P < 0.001, and ***P < 0.0001 compared to the baseline (BL) level prior to stimulus; AVP = arginine vasopressin; FSK = forskolin; Tvp = tolvaptan; Ctrl = control; Ver = verapamil. (c) Vasopressin receptor type-2 (V2R) is localized to primary cilia in renal epithelia, and V2R activation with vasopressin increases adenylyl cyclase (AC) activity. Tolvaptan but not vasopressin elicits a cAMP increase in V2R-knockdown in primary cilium. Unlike tolvaptan which shows cilia-specific cAMP response, AC activator (forskolin) and vasopressin elicit cAMP responses in both cilioplasm and cytoplasm. Cilioplasmic but not cytoplasmic cAMP signaling is repressed below basal levels when cilia bend by fluid-flow. These responses indicate that a cilium may function as a distinct cAMP microdomain, independent from cytoplasmic cAMP.