Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells

Raychowdhury, Malay K.; Ramos, Arnolt J.; Zhang, Peng; McLaughin, Margaret; Dai, Xiao-Qing; Chen, Xing-Zhen; Montalbetti, Nicolás; Cantero, María del Rocío; Ausiello, Dennis A.; Cantiello, Horacio F.

doi:10.1152/ajprenal.90509.2008

Cited by 62 publications

(70 citation statements)

References 43 publications

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“…A link between ciliary Ca 2+ signaling, the polycystin proteins, and cyst development has recently gained attention (42,(46)(47)(48)(49)(50)(51). The fact that the cilia in the InsP3R3 and PC2 knockdown cysts were still present, but absent in InsP3R1 knockdown cysts, by week 8, suggests that cilia may not be essential to the latter stages of cystogenesis and is broadly consistent with recent findings (52).…”

Section: Discussionsupporting

confidence: 81%

Cyst formation following disruption of intracellular calcium signaling

Kuo¹,

DesRochers

Kimmerling

et al. 2014

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

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Mutations in polycystin 1 and 2 (PC1 and PC2) cause the common genetic kidney disorder autosomal dominant polycystic kidney disease (ADPKD). It is unknown how these mutations result in renal cysts, but dysregulation of calcium (Ca 2+ ) signaling is a known consequence of PC2 mutations. PC2 functions as a Ca 2+ -activated Ca 2+ channel of the endoplasmic reticulum. We hypothesize that Ca 2+ signaling through PC2, or other intracellular Ca 2+ channels such as the inositol 1,4,5-trisphosphate receptor (InsP3R), is necessary to maintain renal epithelial cell function and that disruption of the Ca 2+ signaling leads to renal cyst development. The cell line LLC-PK1 has traditionally been used for studying PKD-causing mutations and Ca 2+ signaling in 2D culture systems. We demonstrate that this cell line can be used in long-term (8 wk) 3D tissue culture systems. In 2D systems, knockdown of InsP3R results in decreased Ca 2+ transient signals that are rescued by overexpression of PC2. In 3D systems, knockdown of either PC2 or InsP3R leads to cyst formation, but knockdown of InsP3R type 1 (InsP3R1) generated the largest cysts. InsP3R1 and InsP3R3 are differentially localized in both mouse and human kidney, suggesting that regional disruption of Ca 2+ signaling contributes to cystogenesis. All cysts had intact cilia 2 wk after starting 3D culture, but the cells with InsP3R1 knockdown lost cilia as the cysts grew. Studies combining 2D and 3D cell culture systems will assist in understanding how mutations in PC2 that confer altered Ca 2+ signaling lead to ADPKD cysts. primary cilia | polycysin 2 | calcium release T he commonly occurring genetic kidney disorder, autosomal dominant polycystic kidney disease (ADPKD), is the result of mutations in polycystin 1 or 2 (PC1 or PC2). The progressive cyst formation within all segments of the nephron that defines the disorder leads to renal failure requiring treatment by dialysis and/or organ transplantation (1-3). Altered Ca 2+ signaling is one of several pathways that have been implicated in the disease (4, 5). A major limitation toward elucidating the role of Ca 2+ signaling in cyst formation has been the lack of easily manipulated, physiologically relevant experimental methodologies.In the past, ADPKD research has relied largely upon data from mouse models and cells maintained in 2D cell culture. Mouse models have played a significant role in understanding the biology of cyst formation but are unable to fully recapitulate the physiology of disease progression in humans due to the inherent differences between the species including life span, genetics, and environment. Two-dimensional cell culture has the ability to provide information on signaling pathways and response to therapies in a fast, high-throughput manner, but is incapable of replicating the inherent 3D nature of cyst formation. Advances in 3D tissue culture over the past 2 decades have improved the ability to model cyst development in vitro. However, previously published 3D tissue models of ADPKD have relied upon shortter...

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Section: Discussionsupporting

confidence: 81%

Cyst formation following disruption of intracellular calcium signaling

Kuo¹,

DesRochers

Kimmerling

et al. 2014

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

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“…These findings also confirmed direct phosphorylation by PKA of both PC2 hst and PC2 iv . This is in agreement with previous findings showing that AVP stimulation of a local cAMP pathway activates PC2 in primary cilia of LLC-PK1 renal epithelial cells (23).…”

Section: Discussionsupporting

confidence: 94%

“…Channel open probabilities for the wild type and S812A mutated PC2 showed a bell-shaped dependence on cytoplasmic Ca 2ϩ with a rightward shift in the Ca 2ϩ dependence such that S812A PC2 was 10-fold less sensitive to Ca 2ϩ activation/inactivation than the wild type PC2 channel. Previous immunochemical and electrophysiological studies on primary cilia of LLC-PK1 renal epithelial cells have shown the expression of ciliary vasopressin 2 (V2R) and local cAMP production (23), which regulates ciliary ion channels, including PC2. No information is currently available on the role PKA and its activating patterns play on PC2 channel properties.…”

Section: Polycystin-2 (Pc2) Is a Trp-type Camentioning

confidence: 99%

The cAMP Signaling Pathway and Direct Protein Kinase A Phosphorylation Regulate Polycystin-2 (TRPP2) Channel Function

Cantero

Velázquez

Streets

et al. 2015

Journal of Biological Chemistry

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Background: TRP channels are targets for kinase phosphorylation. Results: PKA phosphorylates and thus controls Polycystin-2 (TRPP2, PC2) channel function. Conclusion: Phosphorylation of Ser-829 in the carboxyl terminus of PC2 is a gating mechanism of channel activity. Significance: Learning how cAMP/PKA local pathways participate in the regulation of PC2 channel function is crucial for understanding renal physiology and the pathogenesis of ADPKD.

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“…Primary cilia have been shown to contain G protein-coupled receptors (GPCR) that signal through cAMP, including receptors for somatostatin and odorants (31). Recently, the type 2 vasopressin receptor (V 2 R) has been identified in primary cilia on renal epithelial cells where it colocalizes with AC5/6 (31).…”

Section: B Cmentioning

confidence: 99%

Polycystin-2 and phosphodiesterase 4C are components of a ciliary A-kinase anchoring protein complex that is disrupted in cystic kidney diseases

Choi¹,

Suzuki²,

Hajarnis³

et al. 2011

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

118

115

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Polycystic kidney disease (PKD) is a genetic disorder that is characterized by cyst formation in kidney tubules. PKD arises from abnormalities of the primary cilium, a sensory organelle located on the cell surface. Here, we show that the primary cilium of renal epithelial cells contains a protein complex comprising adenylyl cyclase 5/6 (AC5/6), A-kinase anchoring protein 150 (AKAP150), and protein kinase A. Loss of primary cilia caused by deletion of Kif3a results in activation of AC5 and increased cAMP levels. Polycystin-2 (PC2), a ciliary calcium channel that is mutated in human PKD, interacts with AC5/6 through its C terminus. Deletion of PC2 increases cAMP levels, which can be corrected by reexpression of wild-type PC2 but not by a mutant lacking calcium channel activity. Phosphodiesterase 4C (PDE4C), which catabolizes cAMP, is also located in renal primary cilia and interacts with the AKAP150 complex. Expression of PDE4C is regulated by the transcription factor hepatocyte nuclear factor-1β (HNF-1β), mutations of which produce kidney cysts. PDE4C is down-regulated and cAMP levels are increased in HNF-1β mutant kidney cells and mice. Collectively, these findings identify PC2 and PDE4C as unique components of an AKAP complex in primary cilia and reveal a common mechanism for dysregulation of cAMP signaling in cystic kidney diseases arising from different gene mutations.

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Vasopressin receptor-mediated functional signaling pathway in primary cilia of renal epithelial cells

Cited by 62 publications

References 43 publications

Cyst formation following disruption of intracellular calcium signaling

Cyst formation following disruption of intracellular calcium signaling

The cAMP Signaling Pathway and Direct Protein Kinase A Phosphorylation Regulate Polycystin-2 (TRPP2) Channel Function

Polycystin-2 and phosphodiesterase 4C are components of a ciliary A-kinase anchoring protein complex that is disrupted in cystic kidney diseases

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