Cellular localization of type 5 and type 6 ACs in collecting duct and regulation of cAMP synthesis

Héliès-Toussaint, Cécile; Aarab, Lotfi; Gasc, Jean-Marie; Verbavatz, Jean‐Marc; Chabardès, Danielle

doi:10.1152/ajprenal.2000.279.1.f185

Cited by 32 publications

(34 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…28 AC5 expression is found in medulla and cortical and outer medullary CD of rats. 16,29,30 We found that expression of AC5 mRNA does not change with variation in fluid intake and AC5 is not up-regulated in AC6 Ϫ/Ϫ mice. Moreover, in preliminary studies, we found that lack of AC5 does not alter basal urine osmolality (WT: 1988 Ϯ 81 and AC5 Ϫ/Ϫ mice: 2089 Ϯ 53 mmol/kg, not significant; n ϭ 4/genotype; mice were kindly provided by P. L. Han 31 ).…”

Section: Discussionmentioning

confidence: 66%

Adenylate Cyclase 6 Determines cAMP Formation and Aquaporin-2 Phosphorylation and Trafficking in Inner Medulla

Rieg

Tang²,

Murray³

et al. 2010

Journal of the American Society of Nephrology

View full text Add to dashboard Cite

Arginine vasopressin (AVP) enhances water reabsorption in the renal collecting duct by vasopressin V 2 receptor (V 2 R)-mediated activation of adenylyl cyclase (AC), cAMP-promoted phosphorylation of aquaporin-2 (AQP2), and increased abundance of AQP2 on the apical membrane. Multiple isoforms of adenylate cyclase exist, and the roles of individual AC isoforms in water homeostasis are not well understood. Here, we found that levels of AC6 mRNA, the most highly expressed AC isoform in the inner medulla, inversely correlate with fluid intake. Moreover, mice lacking AC6 had lower levels of inner medullary cAMP, reduced abundance of phosphorylated AQP2 (at both serine-256 and serine-269), and lower urine osmolality than wild-type mice. Water deprivation or administration of the V 2 R agonist dDAVP did not increase urine osmolality of AC6-deficient mice to the levels of wild-type mice. Furthermore, AC6-deficient mice lacked dDAVP-promoted inner medullary cAMP formation and phosphorylation of serine-269 and had attenuated increases in both phosphorylation of serine-256 and apical membrane AQP2 trafficking. In summary, AC6 expression determines inner medullary cAMP formation and AQP2 phosphorylation and trafficking, the absence of which causes nephrogenic diabetes insipidus. The anti-diuretic hormone arginine-vasopressin (AVP) is the primary regulator of water reabsorption in the renal collecting duct (CD) and is critically involved in the regulation of water balance and maintenance of plasma osmolality. 1 AVP acts on the CD through the G s protein-coupled vasopressin V 2 receptor (V 2 R) to stimulate adenylyl cyclase (AC) and thus the synthesis of cAMP. 2 cAMP activates protein kinase A (PKA), which phosphorylates the water channel aquaporin-2 (AQP2) in its COOH-terminal tail on serine residue 256 (S256), thereby resulting in apical plasma membrane accumulation of AQP2. [3][4][5][6] In addition, cAMP-independent activation of AQP2 has been reported, which may involve V 2 R action of phosphoinositide-specific phospholipase C. 7 Other non-PKA-targeted AQP2 phosphorylation sites include S261, S264, and S269. 8 Phosphorylation of AQP2 at S264 and S269 requires prior phosphorylation of S256. 9 Immunohistochemistry showed that pS269-AQP2 localizes exclusively in the apical plasma

show abstract

Section: Discussionmentioning

confidence: 66%

Adenylate Cyclase 6 Determines cAMP Formation and Aquaporin-2 Phosphorylation and Trafficking in Inner Medulla

Rieg

Tang²,

Murray³

et al. 2010

Journal of the American Society of Nephrology

View full text Add to dashboard Cite

show abstract

“…This isoform is inhibited by Ca 2ϩ in the micromolar range (37) and is not calmodulinsensitive (38). AC6 has been localized in the collecting duct by RT-PCR (29) and in situ hybridization (36). Furthermore, increasing intracellular calcium appears to inhibit AVP-stimulated cAMP in the outer medullary part of the collecting duct (29), supporting the conclusion that AC6 is responsible for vasopressin-dependent cAMP production by outer medullary collecting duct principal cells.…”

Section: Discussionmentioning

confidence: 83%

“…The molecular identity of the adenylyl cyclase responsible for vasopressin-mediated increases in cyclic AMP in IMCD has not been addressed previously, although it has been widely assumed that AC6 is responsible for vasopressin-stimulated cAMP increases because it has been demonstrated to be relatively abundant in collecting duct principal cells (29,36). This isoform is inhibited by Ca 2ϩ in the micromolar range (37) and is not calmodulinsensitive (38).…”

Section: Discussionmentioning

confidence: 99%

Calmodulin Is Required for Vasopressin-stimulated Increase in Cyclic AMP Production in Inner Medullary Collecting Duct

Hoffert

Chou

Fenton

et al. 2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

Calmodulin plays a critical role in regulation of renal collecting duct water permeability by vasopressin. However, specific targets for calmodulin action have not been thoroughly addressed. In the present study, we investigated whether Ca 2؉ /calmodulin regulates adenylyl cyclase activity in the renal inner medullary collecting duct. Rat inner medullary collecting duct suspensions were incubated in the presence or absence of 0.1 nM vasopressin and the calmodulin inhibitors, monodansylcadaverine, W-7, and trifluoperazine, followed by measurement of cAMP. Vasopressin-stimulated cAMP elevation was significantly attenuated in the presence of calmodulin inhibitors. Analysis of transglutaminase 2 knock-out mice confirmed that these compounds were not acting through inhibition of transglutaminase 2 activity. Calmodulin inhibitors also blocked both cholera toxin-and forskolinstimulated cAMP accumulation. In isolated perfused tubules, W-7 reversibly blocked vasopressin-stimulated urea permeability, a process that requires a rise in intracellular cAMP but does not appear to involve protein trafficking to the apical plasma membrane. These results suggest that calmodulin is required for vasopressinstimulated adenylyl cyclase activity in the intact inner medullary collecting duct. Reverse transcription-PCR, immunoblotting, and immunohistochemistry revealed the presence of the calmodulin-sensitive adenylyl cyclase type 3 in the rat collecting duct, an isoform previously not known to be expressed in the collecting duct. Long-term treatment of Brattleboro rats with a vasopressin analog markedly decreased adenylyl cyclase type 3 protein abundance, providing an explanation for long-term down-regulation of vasopressin response in the collecting duct. These studies demonstrate the importance of calmodulin in the regulation of collecting duct adenylyl cyclase activity and transport function.

show abstract

“…Water and urea transport across this epithelium is regulated by the peptide hormone vasopressin via V2-type receptors. Ligand binding triggers an incompletely understood signaling response that includes activation of at least two adenylyl cyclase isoforms via the heterotrimeric G protein G s (7)(8)(9), an increase in intracellular cAMP (10), an increase in cytosolic Ca 2ϩ concentration (11), calmodulin-mediated activation of myosin light-chain kinase (12), and phosphorylation of the water channel aquaporin-2 (AQP2) (13) as well as the urea transporter isoforms A1 and A3 (UT-A1͞3) (14). The work presented here identifies a large number of previously unknown phosphorylation sites in IMCD proteins, including sites in AQP2 and UT-A1͞3.…”

mentioning

confidence: 99%

Quantitative phosphoproteomics of vasopressin-sensitive renal cells: Regulation of aquaporin-2 phosphorylation at two sites

Hoffert

Pisitkun

Wang

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

327

325

View full text Add to dashboard Cite

Protein phosphorylation plays a key role in vasopressin signaling in the renal-collecting duct. Large-scale identification and quantification of phosphorylation events triggered by vasopressin is desirable to gain a comprehensive systems-level understanding of this process. We carried out phosphoproteomic analysis of rat inner medullary collecting duct cells by using a combination of phosphopeptide enrichment by immobilized metal affinity chromatography and phosphorylation site identification by liquid chromatography-mass spectrometry n neutral loss scanning. A total of 714 phosphorylation sites on 223 unique phosphoproteins were identified from inner medullary collecting duct samples treated shortterm with either calyculin A or vasopressin. A number of proteins involved in cytoskeletal reorganization, vesicle trafficking, and transcriptional regulation were identified. Previously unidentified phosphorylation sites were found for membrane proteins essential to collecting duct physiology, including eight sites among aquaporin-2 (AQP2), aquaporin-4, and urea transporter isoforms A1 and A3. Through label-free quantification of phosphopeptides, we identified a number of proteins that significantly changed phosphorylation state in response to short-term vasopressin treatment: AQP2, Bclaf1, LRRC47, Rgl3, and SAFB2. In the presence of vasopressin, AQP2 monophosphorylated at S256 and diphosphorylated AQP2 (pS256͞261) increased in abundance, whereas AQP2 monophosphorylated at S261 decreased, raising the possibility that both sites are involved in vasopressin-dependent AQP2 trafficking. This study reveals the practicality of liquid chromotography-mass spectrometry n neutral loss scanning for large-scale identification and quantification of protein phosphorylation in the analysis of cell signaling in a native mammalian system. LC-MS͞MS ͉ collecting duct ͉ kidney ͉ Collecting Duct Phosphoprotein Database (CDPD) ͉ neutral loss E lucidation of cellular signaling networks requires methodologies for large-scale quantitative phosphoproteomic analysis that can be used to reveal dynamic system-wide changes in protein phosphorylation. Recent studies have introduced two new innovations, namely, immobilized metal affinity chromatography (IMAC) (1-3) and liquid chromatography-mass spectrometry (LC-MS) 3 neutral loss scanning (1-4), to increase the efficiency of phosphopeptide identification. Quantification of phosphopeptides in this setting has been challenging and has been successful so far in cultured cells (5) and yeast (6) but not in native mammalian cells and tissues. Here, we introduce a hybrid approach using IMAC for phosphopeptide enrichment, LC-MS multisequential (LC-MS n ) neutral loss scanning to identify phosphorylated residues in the peptides, and label-free quantification using numerical integration of pseudochromatograms constructed from MS peak heights.The method is used here for analysis of protein phosphorylation in vasopressin-sensitive inner medullary collecting duct (IMCD) cells freshly isolated from rat kidneys. ...

show abstract

Cellular localization of type 5 and type 6 ACs in collecting duct and regulation of cAMP synthesis

Cited by 32 publications

References 31 publications

Adenylate Cyclase 6 Determines cAMP Formation and Aquaporin-2 Phosphorylation and Trafficking in Inner Medulla

Adenylate Cyclase 6 Determines cAMP Formation and Aquaporin-2 Phosphorylation and Trafficking in Inner Medulla

Calmodulin Is Required for Vasopressin-stimulated Increase in Cyclic AMP Production in Inner Medullary Collecting Duct

Quantitative phosphoproteomics of vasopressin-sensitive renal cells: Regulation of aquaporin-2 phosphorylation at two sites

Contact Info

Product

Resources

About