Decreased expression of AQP2 and AQP4 water channels and Na,K-ATPase in kidney collecting duct in AQP3 null mice.
BACKGROUND
Phenotype analysis has demonstrated that AQP3 (aquaporin 3) null mice are polyuric and manifest a urinary concentration defect. In the present study, we report that deletion of AQP3 is also associated with an increased urinary sodium excretion. To investigate further the mechanism of the decreased urinary concentration and significant natriuresis, we examined the segmental and subcellular localization of collecting duct AQPs [AQP2, p-AQP2 (phosphorylated AQP2), AQP3 and AQP4], ENaC (epithelial sodium channel) subunits and Na,K-ATPase by immunoperoxidase and immunofluorescence microscopy in AQP3 null (-/-), heterozygous (+/-) mice, wild-type and unrelated strain of normal mice.


RESULTS
The present study confirms that AQP3 null mice exhibit severe polyuria and polydipsia and demonstrated that they exhibit increased urinary sodium excretion. In AQP3 null mice, there is a marked down-regulation of AQP2 and p-AQP2 both in CNT (connecting tubule) and CCD (cortical collecting duct). Moreover, AQP4 is virtually absent from CNT and CCD in AQP3 null mice. Basolateral AQP2 was virtually absent from AQP3 null mice and normal mice in contrast with rat. Thus the above results demonstrate that no basolateral AQPs are expressed in CNT and CCD of AQP3 null mice. However, in the medullary-collecting ducts, there is no difference in the expression levels and subcellular localization of AQP2, p-AQP2 and AQP4 between AQP3 +/- and AQP3 null mice. Moreover, a striking decrease in the immunolabelling of the alpha1 subunit of Na,K-ATPase was observed in CCD in AQP3 null mice, whereas a medullary-collecting duct exhibited normal labelling. Immunolabelling of all the ENaC subunits in the collecting duct was comparable between the two groups.


CONCLUSIONS
The results improve the possibility that the severe urinary concentrating defect in AQP3 null mice may in part be caused by the decreased expression of AQP2, p-AQP2 and AQP4 in CNT and CCD, whereas the increased urinary sodium excretion may in part be accounted for by Na,K-ATPase in CCD in AQP3 null mice.
Kim
Soo Wan
SW
Water and Salt Research Center, University of Aarhus, DK-8000 Aarhus C, Denmark.
Gresz
Veronika
V
Rojek
Aleksandra
A
Wang
Weidong
W
Verkman
A S
AS
Frøkiaer
Jørgen
J
Nielsen
Søren
S
D000818:Animals; D051399:Aquaporin 2; D051400:Aquaporin 3; D051401:Aquaporin 4; D020346:Aquaporins / Q000096:biosynthesis* / Q000172:deficiency* / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D008817:Mice, Mutant Strains; D009318:Natriuresis / Q000235:genetics; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis* / Q000235:genetics; D014556:Urine
C496024:Aqp2 protein, mouse; C496028:Aqp3 protein, mouse; C496031:Aqp4 protein, mouse; D051399:Aquaporin 2; D051401:Aquaporin 4; D020346:Aquaporins; D051400:Aquaporin 3; C497344:Atp1a1 protein, mouse; D000254:Sodium-Potassium-Exchanging ATPase