Effect of the cGMP pathway on AQP2 expression and translocation: potential implications for nephrogenic diabetes insipidus.
BACKGROUND
Arginine vasopressin (AVP) binding to the V2 receptor (V2R) in renal collecting duct principal cells induces a cAMP signalling cascade resulting in the activation of protein kinase A (PKA), translocation of aquaporin-2 (AQP2) to the apical membrane and an increase in AQP2 expression. Consequently, concentration of urine is initiated. X-linked nephrogenic diabetes insipidus (NDI), characterized by the inability to concentrate urine in response to AVP, is caused by mutations in the V2R gene. Initiation of AQP2 translocation, while circumventing the V2R-cAMP-PKA pathway has been suggested as a putative therapy for these patients. In this respect, the activation of a cAMP-independent and cGMP-dependent pathway for AQP2 membrane insertion by different cyclic guanosine monophosphate (cGMP) pathway activators, such as atrial natriuretic peptide (ANP), l-arginine and 8-bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP), has been put forward. However, it is unclear whether they can increase AQP2 expression.


METHODS
Mouse cortical collecting duct (mpkCCD) cells were incubated with ANP, l-arginine and 8-Br-cGMP for 2 h and subjected to immunocytochemistry and cell surface biotinylation assays to examine their effect on AQP2 translocation. To test the effect of cGMP pathway activators on AQP2 expression, the mpkCCD cells were treated with dDAVP, ANP and l-arginine for 4 days, or with 8-Br-cGMP for the last day. AQP2 protein levels were determined by immunoblotting.


RESULTS
ANP, l-arginine and 8-Br-cGMP induced the translocation of AQP2 in the mpkCCD cells. However, in contrast to dDAVP, ANP, l-arginine and 8-Br-cGMP did not increase the expression of AQP2.


CONCLUSIONS
Our results suggest that while activators of the cGMP pathway are likely beneficial in the treatment of X-linked NDI, their ability to relieve NDI in the patients may be improved when combined with agents stimulating AQP2 expression.
Boone
Michelle
M
Department of Physiology, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands. p.deen@ncmls.ru.nl
Kortenoeven
Marleen
M
Robben
Joris H
JH
Deen
Peter M T
PM
D000818:Animals; D065092:Antidiuretic Hormone Receptor Antagonists; D051399:Aquaporin 2 / Q000378:metabolism*; D001120:Arginine / Q000494:pharmacology; D001127:Arginine Vasopressin / Q000378:metabolism; D009320:Atrial Natriuretic Factor / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology / Q000502:physiology*; D018500:Diabetes Insipidus, Nephrogenic / Q000188:drug therapy*; D006801:Humans; D007685:Kidney Tubules, Collecting / Q000378:metabolism* / Q000473:pathology; D008297:Male; D023421:Models, Animal; D009025:Morpholines / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000502:physiology*; D017483:Receptors, Vasopressin / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013141:Spiro Compounds / Q000494:pharmacology
D065092:Antidiuretic Hormone Receptor Antagonists; C496024:Aqp2 protein, mouse; D051399:Aquaporin 2; D009025:Morpholines; D017483:Receptors, Vasopressin; D013141:Spiro Compounds; D001127:Arginine Vasopressin; C016276:8-bromocyclic GMP; D009320:Atrial Natriuretic Factor; D001120:Arginine; C103281:satavaptan; D006152:Cyclic GMP