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1 Laboratory of Kidney and Electrolyte Metabolism, National Institutes of Health, Bethesda, Maryland 20892-1603; and 2 Departments of Medicine and Physiology, Cardiovascular Research Institute, University of California, San Francisco, California 94143-0521
Aquaporin (AQP)-3 and AQP4 water channels are expressed at the
basolateral membrane of mammalian collecting duct epithelium. To
determine the contribution of AQP4 to water permeability in the initial
inner medullary collecting duct (IMCD), osmotic water permeability
(Pf) was
compared in isolated perfused IMCD segments from wild-type and AQP4
knockout mice. The AQP4 knockout mice were previously found to have
normal gross appearance, survival, growth, and kidney morphology and a
mild urinary concentrating defect (T. Ma, B. Yang, A. Gillespie, E. J. Carlson, C. J. Epstein, and A. S. Verkman. J. Clin.
Invest. 100: 957-962, 1997).
Transepithelial Pf was measured
in microdissected IMCDs after 18-48 h of water deprivation and in
the presence of 0.1 nM arginine vasopressin (to make basolateral
Pf rate
limiting). Pf
values (37°C; means ± SE in cm/s × 10
3) were 56.0 ± 8.5 for wild-type mice (n = 5) and 13.1 ± 3.7 for knockout mice (n = 6)
(P < 0.001). Northern blot analysis
of kidney showed that transcript expression of AQP1, AQP2, AQP3, and
AQP6 were not affected by AQP4 deletion. Immunoblot analysis indicated no differences in protein expression of AQP1, AQP2, or AQP3, and immunoperoxidase showed no differences in staining patterns.
Coexpression of AQP3 and AQP4 in Xenopus
laevis oocytes showed additive water permeabilities,
suggesting that AQP4 deletion does not affect AQP3 function. These
results indicate that AQP4 is responsible for the majority of
basolateral membrane water movement in IMCD but that its deletion is
associated with a very mild defect in urinary concentrating ability.
kidney; urinary concentration; transgenic mice; vasopressin
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