Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Lykke K
,
Assentoft M
,
Fenton RA
,
Rosenkilde MM
,
MacAulay N
.
???displayArticle.abstract???
Herein, we investigated whether G protein-coupled signaling via the vasopressin receptors of the V1a and V2 subtypes (V1aR and V2R) could be obtained as a direct response to hyperosmolar challenges and/or whether hyperosmolar challenges could augment classical vasopressin-dependent V1aR signaling. The V1aR-dependent response was monitored indirectly via its effects on aquaporin 4 (AQP4) when heterologously expressed in Xenopus oocytes and V1aR and V2R function was directly monitored following heterologous expression in COS-7 cells. A tendency toward an osmotically induced, V1aR-mediated reduction in AQP4-dependent water permeability was observed, although osmotic challenges failed to mimic vasopressin-dependent V1aR-mediated internalization of AQP4. Direct monitoring of inositol phosphate (IP) production of V1aR-expressing COS-7 cells demonstrated an efficient vasopressin-dependent response that was, however, independent of hyperosmotic challenges. Similarly, the cAMP production by the V2R was unaffected by hyperosmotic challenges although, in contrast to the V1aR, the V2R displayed an ability to support alternative signaling (IP production) at higher concentration of vasopressin. V1aR and V2R respond directly to vasopressin exposure, but they do not have an ability to act as osmo- or volume sensors when exposed to an osmotic gradient in the absence or presence of vasopressin.
Figure 1. V1aR-dependent downregulation of AQP4. (A) Volume traces obtained from an uninjected oocyte (left panel) and an AQP4/V1aR-expressing oocyte challenged with an osmotic gradient of 50 mOsm mannitol for 30 sec. (B) Relative water permeability of oocytes expressing AQP4 (open circles; n = 5) or coexpressing AQP4/V1aR (filled circles, n = 20) exposed to 1 μmol/L vasopressin as marked by the black bar. (C) Relative water permeability of oocytes expressing AQP4 (open circles; n = 6) or coexpressing AQP4/V1aR (filled circles; n = 22) when exposed to repeated osmotic challenges. (D) Relative water permeability of oocytes coexpressing AQP4/mGluR1a and exposed to 500 μmol/L glutamate as indicated by the black bar (filled symbols, n = 8) or kept in control solution (open symbols, n = 8, not exposed to glutamate). The groups were compared with two-way analysis of variance (ANOVA) with Å Ãdákâs multiple comparison post hoc test. *P < 0.05.
Figure 2. V1aR-dependent internalization of AQP4. (A) Confocal laser scanning microscopy of oocytes expressing either AQP4 (left panel) or AQP4/V1aR (right panel) immune-labeled for AQP4. The upper panels are representative images of oocytes exposed to control solution without vasopressin for 80 min. The middle panels are representative images of oocytes kept in control solution for 20 min and then treated with 1 μmol/L vasopressin for 60 min. The lower panels are representative images of oocytes treated with a 50 mOsm hyperosmolar gradient for 30 sec every 10 min of an 80-min incubation period. (B) Oocyte plasma membrane fluorescence intensity normalized to that of the oocytes kept in control solution, n = 5 experiments with 3â6 oocytes per condition. The indicated groups were compared using one-way analysis of variance (ANOVA) with Å Ãdákâs multiple comparison post hoc test. *P < 0.05; ns, not significant.
Figure 3. Hyperosmolar effects on V1aR-dependent inositol phosphate (IP) production. (A) Vasopressin doseâresponse curve measured with IP production in COS-7 cells transfected with either vector alone or with V1aR, n = 7. (B) IP production of COS-7 cells transfected with either vector alone or with V1aR upon addition of vasopressin (1 μmol/L) or various hyperosmolar treatments as indicated on graph, n = 3â8. (C) Effect of hyperosmolar treatment (50 or 150 mOsm) on the vasopressin doseâresponse curve, n = 3â8. (D) cAMP production obtained in COS-7 cells transfected with either vector alone or with V1aR in response to vasopressin exposure (1 μmol/L) or various hyperosmolar treatments as indicated on graph, n = 4â6. Data obtained with V1aR-transfected cells were compared to those obtained with vector-transfected cells using two-way analysis of variance (ANOVA) with Å Ãdákâs multiple comparison post hoc test and indicated with * (note: the difference between the vasopressin-induced cAMP production in vector- and V1aR-transfected cells was not statistically significant even when compared with Studentâs t-test). Statistical significance of vasopressin-induced cAMP production in vector-transfected cells was determined with two-way ANOVA with a Dunnettâs multiple comparison test and indicated with #. #P < 0.05; ***P < 0.001; ns, not significant.
Figure 4. Hyperosmolar effects on V2R-dependent cAMP and inositol phosphate (IP) production. (A) Vasopressin doseâresponse curve measured with cAMP response of COS-7 cells transfected with either vector alone or with V2R, n = 5. (B) cAMP production of COS-7 cells transfected with either vector alone or with V2R upon addition of vasopressin (1 μmol/L) or various hyperosmolar treatments as indicated on graph, n = 4â6. (C) IP production (black and gray circles) and cAMP accumulation (dashed line, adapted from panel A) of COS-7 cells transfected with V2R upon vasopressin exposure, n = 4â7. Data obtained with V2R-transfected cells were compared to those obtained with vector-transfected cells using two-way ANOVA with Å Ãdákâs multiple comparison post hoc test and indicated with *. Statistical significance of vasopressin-induced cAMP production in vector-transfected cells was determined with two-way ANOVA with a Dunnettâs multiple comparison test and indicated with #. #P < 0.05; ***P < 0.001; ns, not significant.
Ancellin,
Homologous and heterologous acute desensitization of vasopressin V1a receptor in Xenopus oocytes.
1998, Pubmed,
Xenbase
Ancellin,
Homologous and heterologous acute desensitization of vasopressin V1a receptor in Xenopus oocytes.
1998,
Pubmed
,
Xenbase
Bourque,
Central mechanisms of osmosensation and systemic osmoregulation.
2008,
Pubmed
Briley,
The cloned vasopressin V1a receptor stimulates phospholipase A2, phospholipase C, and phospholipase D through activation of receptor-operated calcium channels.
1994,
Pubmed
Chen,
Vasopressin induction of long-lasting potentiation of synaptic transmission in the dentate gyrus.
1993,
Pubmed
Ciura,
Hypertonicity sensing in organum vasculosum lamina terminalis neurons: a mechanical process involving TRPV1 but not TRPV4.
2011,
Pubmed
Ciura,
Transient receptor potential vanilloid 1 is required for intrinsic osmoreception in organum vasculosum lamina terminalis neurons and for normal thirst responses to systemic hyperosmolality.
2006,
Pubmed
Fenton,
Differential water permeability and regulation of three aquaporin 4 isoforms.
2010,
Pubmed
,
Xenbase
Graham,
A new technique for the assay of infectivity of human adenovirus 5 DNA.
1973,
Pubmed
Grunnet,
KCNQ1 channels sense small changes in cell volume.
2003,
Pubmed
,
Xenbase
Izumi,
Regulation of V2R transcription by hypertonicity and V1aR-V2R signal interaction.
2008,
Pubmed
Jójárt,
Immunoelectronhistochemical evidence for innervation of brain microvessels by vasopressin-immunoreactive neurons in the rat.
1984,
Pubmed
Juul,
The physiological and pathophysiological functions of renal and extrarenal vasopressin V2 receptors.
2014,
Pubmed
Kissow,
Glucagon-like peptide-1 (GLP-1) receptor agonism or DPP-4 inhibition does not accelerate neoplasia in carcinogen treated mice.
2012,
Pubmed
Kleindienst,
Protective effect of the V1a receptor antagonist SR49059 on brain edema formation following middle cerebral artery occlusion in the rat.
2006,
Pubmed
Kortenoeven,
Renal aquaporins and water balance disorders.
2014,
Pubmed
Krämer,
Regulative interactions of the osmosensing C-terminal domain in the trimeric glycine betaine transporter BetP from Corynebacterium glutamicum.
2009,
Pubmed
Kudo,
Effect of peritubular hypertonicity on water and urea transport of inner medullary collecting duct.
1992,
Pubmed
Landgraf,
Central release of vasopressin: stimuli, dynamics, consequences.
1992,
Pubmed
Liu,
Arginine-vasopressin V1 but not V2 receptor antagonism modulates infarct volume, brain water content, and aquaporin-4 expression following experimental stroke.
2010,
Pubmed
Lolait,
Cloning and characterization of a vasopressin V2 receptor and possible link to nephrogenic diabetes insipidus.
1992,
Pubmed
Lolait,
Molecular biology of vasopressin receptors.
1995,
Pubmed
Moeller,
Vasopressin-dependent short-term regulation of aquaporin 4 expressed in Xenopus oocytes.
2009,
Pubmed
,
Xenbase
Nadler,
Effects of hypertonicity on ADH-stimulated water permeability in rat inner medullary collecting duct.
1990,
Pubmed
Nathanson,
Mechanisms of subcellular cytosolic Ca2+ signaling evoked by stimulation of the vasopressin V1a receptor.
1992,
Pubmed
,
Xenbase
Plant,
TRPs in mechanosensing and volume regulation.
2014,
Pubmed
Reymond-Marron,
The vasopressin-induced excitation of hypoglossal and facial motoneurons in young rats is mediated by V1a but not V1b receptors, and is independent of intracellular calcium signalling.
2006,
Pubmed
Steen,
Biased and g protein-independent signaling of chemokine receptors.
2014,
Pubmed
Szmydynger-Chodobska,
Increased expression of vasopressin v1a receptors after traumatic brain injury.
2004,
Pubmed
Terrillon,
Heterodimerization of V1a and V2 vasopressin receptors determines the interaction with beta-arrestin and their trafficking patterns.
2004,
Pubmed
Thibonnier,
Molecular cloning, sequencing, and functional expression of a cDNA encoding the human V1a vasopressin receptor.
1994,
Pubmed
van Leeuwen,
Quantitative light microscopic autoradiographic localization of binding sites labelled with [3H]vasopressin antagonist d(CH2)5Tyr(Me)VP in the rat brain, pituitary and kidney.
1987,
Pubmed
Violin,
Beta-arrestin-biased ligands at seven-transmembrane receptors.
2007,
Pubmed
Wrobel,
Excitatory action of vasopressin in the brain of the rat: role of cAMP signaling.
2011,
Pubmed
Zampighi,
A method for determining the unitary functional capacity of cloned channels and transporters expressed in Xenopus laevis oocytes.
1995,
Pubmed
,
Xenbase
Zeuthen,
Water transport by Na+-coupled cotransporters of glucose (SGLT1) and of iodide (NIS). The dependence of substrate size studied at high resolution.
2006,
Pubmed
,
Xenbase
Zhao,
Vasopressin-induced cytoplasmic and nuclear calcium signaling in embryonic cortical astrocytes: dynamics of calcium and calcium-dependent kinase translocation.
2003,
Pubmed
Zhu,
Dual signaling potential is common among Gs-coupled receptors and dependent on receptor density.
1994,
Pubmed
