Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. 1994 Jul 5; 91(14): 6269–6273.

Molecular cloning and expression of a member of the aquaporin family with permeability to glycerol and urea in addition to water expressed at the basolateral membrane of kidney collecting duct cells.


Water transport in highly water-permeable membranes is conducted by water-selective pores--namely, water channels. The recent cloning of water channels revealed the water-selective characteristics of these proteins when expressed in Xenopus oocytes or reconstituted in liposomes. Currently, it is assumed that the function of water channels is to transport only water. We now report the cloning of a member of the water channel that also transports nonionic small molecules such as urea and glycerol. We named this channel aquaporin 3 (AQP3) for its predominant water permeability. AQP3 has amino acid sequence identity with major intrinsic protein (MIP) family proteins including AQP-channel-forming integral membrane protein, AQP-collecting duct, MIP, AQP-gamma tonoplast intrinsic protein, nodulin 26, and glycerol facilitator (33-42%). Thus, AQP3 is an additional member of the MIP family. Osmotic water permeability of Xenopus oocytes measured by videomicroscopy was 10-fold higher in oocytes injected with AQP3 transcript than with water-injected oocytes. The increase in osmotic water permeability was inhibited by HgCl2, and this effect was reversed by a reducing agent, 2-mercaptoethanol. Although to a smaller degree, AQP3 also facilitated the transport of nonionic small solutes such as urea and glycerol, while the previously cloned water channels are permeable only to water when expressed in Xenopus oocytes. AQP3 mRNA was expressed abundantly in kidney medulla and colon. In kidney, it was exclusively immunolocalized at the basolateral membrane of collecting duct cells. AQP3 may function as a water and urea exit mechanism in antidiuresis in collecting duct cells.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.2M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Preston GM, Carroll TP, Guggino WB, Agre P. Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein. Science. 1992 Apr 17;256(5055):385–387. [PubMed]
  • Fushimi K, Uchida S, Hara Y, Hirata Y, Marumo F, Sasaki S. Cloning and expression of apical membrane water channel of rat kidney collecting tubule. Nature. 1993 Feb 11;361(6412):549–552. [PubMed]
  • Maurel C, Reizer J, Schroeder JI, Chrispeels MJ. The vacuolar membrane protein gamma-TIP creates water specific channels in Xenopus oocytes. EMBO J. 1993 Jun;12(6):2241–2247. [PMC free article] [PubMed]
  • Preston GM, Agre P. Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodaltons: member of an ancient channel family. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11110–11114. [PMC free article] [PubMed]
  • Lanahan A, Williams JB, Sanders LK, Nathans D. Growth factor-induced delayed early response genes. Mol Cell Biol. 1992 Sep;12(9):3919–3929. [PMC free article] [PubMed]
  • Hasegawa H, Zhang R, Dohrman A, Verkman AS. Tissue-specific expression of mRNA encoding rat kidney water channel CHIP28k by in situ hybridization. Am J Physiol. 1993 Jan;264(1 Pt 1):C237–C245. [PubMed]
  • Nielsen S, Smith BL, Christensen EI, Agre P. Distribution of the aquaporin CHIP in secretory and resorptive epithelia and capillary endothelia. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7275–7279. [PMC free article] [PubMed]
  • Bondy C, Chin E, Smith BL, Preston GM, Agre P. Developmental gene expression and tissue distribution of the CHIP28 water-channel protein. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4500–4504. [PMC free article] [PubMed]
  • Höfte H, Hubbard L, Reizer J, Ludevid D, Herman EM, Chrispeels MJ. Vegetative and Seed-Specific Forms of Tonoplast Intrinsic Protein in the Vacuolar Membrane of Arabidopsis thaliana. Plant Physiol. 1992 Jun;99(2):561–570. [PMC free article] [PubMed]
  • Baker ME, Saier MH., Jr A common ancestor for bovine lens fiber major intrinsic protein, soybean nodulin-26 protein, and E. coli glycerol facilitator. Cell. 1990 Jan 26;60(2):185–186. [PubMed]
  • Wistow GJ, Pisano MM, Chepelinsky AB. Tandem sequence repeats in transmembrane channel proteins. Trends Biochem Sci. 1991 May;16(5):170–171. [PubMed]
  • Pao GM, Wu LF, Johnson KD, Höfte H, Chrispeels MJ, Sweet G, Sandal NN, Saier MH., Jr Evolution of the MIP family of integral membrane transport proteins. Mol Microbiol. 1991 Jan;5(1):33–37. [PubMed]
  • Macey RI. Transport of water and urea in red blood cells. Am J Physiol. 1984 Mar;246(3 Pt 1):C195–C203. [PubMed]
  • Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. [PubMed]
  • Zhang RB, Logee KA, Verkman AS. Expression of mRNA coding for kidney and red cell water channels in Xenopus oocytes. J Biol Chem. 1990 Sep 15;265(26):15375–15378. [PubMed]
  • Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. [PMC free article] [PubMed]
  • Muramatsu S, Mizuno T. Nucleotide sequence of the region encompassing the glpKF operon and its upstream region containing a bent DNA sequence of Escherichia coli. Nucleic Acids Res. 1989 Jun 12;17(11):4378–4378. [PMC free article] [PubMed]
  • Weissenborn DL, Wittekindt N, Larson TJ. Structure and regulation of the glpFK operon encoding glycerol diffusion facilitator and glycerol kinase of Escherichia coli K-12. J Biol Chem. 1992 Mar 25;267(9):6122–6131. [PubMed]
  • Gorin MB, Yancey SB, Cline J, Revel JP, Horwitz J. The major intrinsic protein (MIP) of the bovine lens fiber membrane: characterization and structure based on cDNA cloning. Cell. 1984 Nov;39(1):49–59. [PubMed]
  • Sandal NN, Marcker KA. Soybean nodulin 26 is homologous to the major intrinsic protein of the bovine lens fiber membrane. Nucleic Acids Res. 1988 Oct 11;16(19):9347–9347. [PMC free article] [PubMed]
  • Yamamoto YT, Cheng CL, Conkling MA. Root-specific genes from tobacco and Arabidopsis homologous to an evolutionarily conserved gene family of membrane channel proteins. Nucleic Acids Res. 1990 Dec 25;18(24):7449–7449. [PMC free article] [PubMed]
  • Rao Y, Jan LY, Jan YN. Similarity of the product of the Drosophila neurogenic gene big brain to transmembrane channel proteins. Nature. 1990 May 10;345(6271):163–167. [PubMed]
  • Verkman AS. Water channels in cell membranes. Annu Rev Physiol. 1992;54:97–108. [PubMed]
  • Alemohammad MM, Knowles CJ. Osmotically induced volume and turbidity changes of Escherichia coli due to salts, sucrose and glycerol, with particular reference to the rapid permeation of glycerol into the cell. J Gen Microbiol. 1974 May;82(1):125–142. [PubMed]
  • Heller KB, Lin EC, Wilson TH. Substrate specificity and transport properties of the glycerol facilitator of Escherichia coli. J Bacteriol. 1980 Oct;144(1):274–278. [PMC free article] [PubMed]
  • You G, Smith CP, Kanai Y, Lee WS, Stelzner M, Hediger MA. Cloning and characterization of the vasopressin-regulated urea transporter. Nature. 1993 Oct 28;365(6449):844–847. [PubMed]
  • Chou CL, Knepper MA. Inhibition of urea transport in inner medullary collecting duct by phloretin and urea analogues. Am J Physiol. 1989 Sep;257(3 Pt 2):F359–F365. [PubMed]
  • Agre P, Sasaki S, Chrispeels MJ. Aquaporins: a family of water channel proteins. Am J Physiol. 1993 Sep;265(3 Pt 2):F461–F461. [PubMed]
  • Fischbarg J, Kuang KY, Vera JC, Arant S, Silverstein SC, Loike J, Rosen OM. Glucose transporters serve as water channels. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3244–3247. [PMC free article] [PubMed]
  • Zhang R, Alper SL, Thorens B, Verkman AS. Evidence from oocyte expression that the erythrocyte water channel is distinct from band 3 and the glucose transporter. J Clin Invest. 1991 Nov;88(5):1553–1558. [PMC free article] [PubMed]
  • Hasegawa H, Skach W, Baker O, Calayag MC, Lingappa V, Verkman AS. A multifunctional aqueous channel formed by CFTR. Science. 1992 Nov 27;258(5087):1477–1479. [PubMed]
  • Strange K, Spring KR. Cell membrane water permeability of rabbit cortical collecting duct. J Membr Biol. 1987;96(1):27–43. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...