This Article Abstract Full Text (PDF) Submit a related Letter to the Editor Purchase Article View Shopping Cart Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Inaba, S. Articles by Miyamori, I. Search for Related Content PubMed PubMed Citation Articles by Inaba, S. Articles by Miyamori, I.

The Property of a Novel V2 Receptor Mutant in a Patient with Nephrogenic Diabetes Insipidus

Third Department of Internal Medicine, Fukui Medical University, Fukui, Japan

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
Nephrogenic diabetes insipidus (NDI) is characterized by resistance of the kidneys to the action of arginine vasopressin (AVP); X-linked recessive NDI is caused by an inactivating mutation of the vasopressin type-2 (V2) receptor. Several missense mutations in the first or second extracellular loop of the V2 receptor have been reported, and some of these mutant receptors were confirmed to have reduced affinities for ligand binding. We detected a novel V2 receptor gene mutation, a substitution of cysteine for arginine-104 (R104C) located in the first extracellular loop of the V2 receptor, in a patient with congenital NDI. Functional analysis by transient expression studies with COS-7 cells showed binding capacity of R104C mutant diminished as 10% of wild type, but binding affinity was strong rather than wild type. In the result of AVP stimulation studies, maximum cAMP accumulation of R104C decreased as 50% of wild type. On the other hand, a designed mutant receptor, substituted serine for arginine-104 as a model of modified R104C mutant receptor removed the influence of the sulfhydryl group in cysteine-104, recovered binding capacity up to 50% of wild type and maximum cAMP accumulation as 82% of wild type. Our study demonstrated that the R104C mutation of the V2 receptor was a cause of NDI. The mechanism of renal resistance to AVP was the reduction of ligand binding, and adenylyl cyclase activation depended on the V2 receptor. In addition, we confirmed that the sulfhydryl group of the cysteine-104 caused most part of R104C mutant receptor dysfunction.

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
CONGENITAL NEPHROGENIC diabetic insipidus (NDI) is a hereditary disorder characterized by a diminished renal response to arginine vasopressin (AVP). Patients with NDI are unable to produce concentrated urine in response to the administration of AVP and, thus, display polyuria and polydipsia as symptoms. Most cases of congenital NDI are inherited in an X-linked recessive manner; this form of NDI has been mapped to the q28-qter position of the human X chromosome by genetic linkage (1, 2). Following the molecular cloning of human vasopressin type-2 (V2) receptor complementary DNA (cDNA), genomic sequences have been identified and characterized (3, 4, 5). The sequences of the cDNA indicate coding for a protein with seven hydrophobic segments, suggesting the receptors belongs to the superfamily of seven-transmembrane-domain receptors that transduce hormonal signals coupling with G proteins. It is now well established that inactivating mutations of the gene cause most cases of X-linked recessive NDI. More than 70 different mutations have been identified; the great majorities are missense and nonsense mutations. Furthermore, 18 frameshift mutations due to nucleotide deletions or insertions (up to 35 bp) and four large deletions have been reported.

Functional characterization of missense mutations revealed three different molecular mechanisms that cause: 1) low ligand-binding affinities; 2) diminished coupling with G protein and subsequent activation; and 3) obstruction of transport. In the first or second extracellular loop, most of the missense mutations substitute an extra cysteine and some of them were confirmed as a cause of reduction in ligand-binding affinity (6, 7). Exchanged cysteine forming a false disulfide bond with another cysteine that would otherwise form another functional bond, and it has been believed that the structural changes of the disulfide bonds reduce the receptor-binding affinity. However, it is suggested that the binding sites of vasopressin V1a receptor was located in the transmembrane domain, and the same agonist-binding site was shared by all members of this receptor family (8). Therefore, it is unclear how these mutations in the first or second extracellular domain reduce its ligand-binding affinities. Besides, they have never been clarified the influence of substituted cysteine on the receptor functions.

In this study, we conducted direct DNA sequencing of the V2 receptor gene from a Japanese patient with congenital NDI and found a novel V2 receptor point mutation from arginine 104 to cysteine in the first extracellular loop. To understand the mechanism of renal resistance to AVP, we expressed the mutant receptor in COS-7 cells to characterize its functional properties.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Patient profile

A 53-yr-old male patient had a history of polyuria and polydipsia from infancy. His daily urine output was typically 7 or 8 L and showed low osmotic pressure despite high plasma AVP concentration (10.9 pg/mL). A water deprivation test showed an inability to produce concentrated urine. At the end of the test, urinary osmotic pressure was 293 mOsm/kg and the serum osmotic pressure was 297 mOsm/kg; urinary osmotic pressure scarcely rose in response to AVP administration. Several of the patient’s male relatives, traced back for several generations, had a similar history of polyuria and polydipsia (Fig. 1).

View larger version (15K): [in this window] [in a new window]   Figure 1. Pedigree of NDI relatives. , Women; , men; Roman numerals, the generation; circles with a central dot, obligatory carriers; , affected subjects. A slash (/) through a symbol indicates that the subject is deceased. The proband is shown by an arrow.

Genomic V2 receptor DNA amplification and sequencing

Genomic DNA was extracted from peripheral white blood cells with a DNA isolation kit (QIAamp Blood Kit; QIAGEN, Dusseldorf, Germany). Three coding exons of the V2 receptor gene were amplified by PCR with oligonucleotide primers: (1F: 5'-AGTCCGCACATCACCTCCA-3', 2F: 5'-CTGTGCCTGGGCATCCCTCT-3', 3F: 5'-TGGCTCTGTTCCAAGTGCTG-3', 4F: 5'-TGACTGCTGGGCCTGCTTTG-3', 5F: 5'-TCTATGTGCTGTGCTGGGCA-3', 1R: 5'-TGTCCAGTGGCCTCTCCTG-3', 2R: 5'-CCATCTGCAGATACTTCACG-3', 3R: 5'-GTAGGTGCCACGAACACCAT-3', 4R: 5'-CTTCCAGAGGTGCCTCCGGG-3', 5R: 5'-CTGAGCTTCTCAAAGCCTCT-3'). Amplification was performed with Taq polymerase according to the recommendation of Perkin-Elmer Corp. (Norwalk, CT). Amplification was carried out at 94 C for 3 min and then for 30 cycles at the following settings: 94 C for 1 min, 60 C for 1 min, 72 C for 2 min. Amplified products were purified on 1.5% agarose gels before sequencing.

Direct sequencing was performed by the dideoxy method using a sequencing system (Thermo Sequenase pre-mixed cycle sequencing kit; Amersham Pharmacia Biotech, Sunnyvale, CA) and an automatic DNA sequencer (SQ-5500; Hitachi, Tokyo, Japan).

Isolation of wild-type human V2 receptor cDNA

Human kidney RNA was extracted from a kidney surgically removed from a patient with renal cell carcinoma. Extraction was performed with the use of a RNA extraction kit (Rneasy Mini Kit; QIAGEN). Synthesis of cDNA was performed with an avian myeloblastosis virus reverse transcriptase (Takara, Otsu, Japan). All steps were performed in duplicate, starting with the reverse transcriptase reaction. The solution was incubated at 42 C for 30 min with random primer (Takara). The full-length V2 receptor cDNA was amplified by PCR.

Site-directed mutagenesis

The full-length V2 receptor cDNA was cloned into the pKF18k plasmid (Takara). Positive clones were completely sequenced by the dideoxy method. Site-directed mutagenesis was performed using the Mutan-Super Express kit (Takara) with two mutagenesis primers (M1: 5'-GCCACCGACTGCTTCCGTG-3' and M2: 5'-GCCACCGACAGCTTCCGTG-3', corresponding to nucleotides +301 to +319, sequence numbering according to GenBank entry Z11687) phosphorylated at the 5' end. These primers produce single base mutations: the M1 primer converts the arginine (CGC)-104 codon to a cysteine (TGC) to produce a R104C mutant and the M2 primer converts the same codon to serine (AGC) to produce a R104S mutant. The substituted sequences were checked by the direct sequencing method. These three V2 receptor cDNAs were amplified by PCR with the following primers: sense 5'-CTTGCTCCTCAGGCAGAGGC-3' and antisense 5'-TCTAGAGGCAAGACACCCAAC-3' (corresponding to nucleotides -71 to -52 and +1123 to +1143). They were then introduced in a sense orientation into the expression plasmid pTargeT (Promega Corp., Madison, WI).

Transient expression in mammalian cells

We cultured COS-7 cells (JCRB9127; Health Science Research Resources Bank, Osaka, Japan) in DMEM (Nippon Paramecia Corp., Tokyo, Japan) with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin at an atmosphere of 5% CO₂ and 95% O₂ at 37 C. The cells were plated in a 100-mm dish in a subconfluent and transfected by the diethylaminoethyl-dextran method (9). Plasmid constructs (10 µg) were mixed with 3.5 mg/mL diethylaminoethyl-dextran in 10 mM Tris-HCl (pH 7.5) in a final volume of 180 µL and incubated at room temperature for 30 min. The plasmid mixture was diluted with 4 mL DMEM with 10 mM Tris-HCl (pH 7.5) and applied to the cells, which had already had the growth medium removed and had been rinsed with Dulbecco’s phosphate-buffered saline (D-PBS) twice. After incubation for 4 h at 37 C, the cells were washed twice with D-PBS, exposed to 100 µL chloroquine in DMEM with 10 mM Tris-HCl (pH 7.5), and incubated at 37 C for 3 h. The cells were again washed twice with D-PBS, returned to fresh growth medium, and incubated for 48 h at 37 C. The transfected cells were assayed for radioligand binding and adenylyl cyclase activity.

To check transfection efficiencies, we determined the amounts of messenger RNA (mRNA) by quantitative RT-PCR analysis. Total RNA of 48 h after transfected cells were obtained with Rneasy Mini Kit (QIAGEN). Reverse transcriptase reactions were performed with each 1 µg total RNA using a RNA PCR Kit (Takara), and produced receptor cDNA were amplified and determined these quantity by Light Cycler (Roche Molecular Biochemicals, Mannheim, West Germany) with Light Cycler DNA Master SYBR Green I (Roche), primers 1F and 5R.

Binding assay

The AVP binding assay was performed 24 h after the cells were plated. After a 24-h transfection, the cells were plated in 12-well plates at a density of 3.0 x 10⁵ cells per well. Each well was washed twice with ice-cold D-PBS, then filled with 0.5 mL DMEM with 1% BSA containing the appropriate dilutions of [³H]-AVP ([phenylalanyl-3,4,5-³H(N)]-Vasopressin (75 Ci/mmol; NEN Life Science Products, Tokyo, Japan). The samples were incubated on ice for 2 h. Nonspecific binding was determined in the presence of 10 µM unlabeled AVP under the same conditions. At the end of the incubation, the wells were washed twice with ice-cold D-PBS. Then, 0.5 mL 0.1 N NaOH was added to extract the bound radioligand. After incubation for 30 min at 37 C, fluid from the wells was transferred to scintillation vials containing 5 mL aqueous counting scintillant (ACSII; Amersham Pharmacia Biotech, Arlington Heights, IL), and radioactivity was measured by a liquid scintillation counter (LSC-3500; Aloka, Tokyo, Japan). The number of cells per well was determined by trypsinization and counting of cells in five wells; the average cell number was used to calculate sites per cell. The data from radioligand binding studies were analyzed by a computer program, GraphPad PRISM version 2.1 (GraphPad Software, Inc., San Diego, CA).

Adenylyl cyclase assay

The cAMP assay was performed 24 h after the cells were plated. After a 24 h transfection, the cells were plated in 24-well plates at a density of 1.5 x 10⁵ cells per well. Each well was washed twice with D-PBS, then filled with 0.5 mL DMEM with 1% BSA and 0.5 mM 1-isobutyl-3-methylxamine containing the appropriate dilutions of AVP. After incubation for 10 min at 37 C, the reactions were stopped by immediate removal of the medium. We determined the quantity of intracellular cAMP with the cAMP extraction and detection kits (cAMP EIA System; Amersham Pharmacia Biotech). The data from cAMP assays were analyzed with GraphPad PRISM version 2.1.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
Sequencing of the V2 receptor gene

Direct sequencing of the V2 receptor gene in the patient with NDI identified a C to T transition at nucleotide position +310 in exon 2 (Fig. 2). The mutation leads to a predicted change of arginine (codon 104, CGC) to a cysteine (TGC). This is a novel missense mutation encoding within the first extracellular loop of the V2 receptor polypeptide, named R104C (Fig. 3). An A to G mutation at nucleotide +927 was also identified in this patient (data not shown). This mutation was previously described by Pan et al. (10) and leads to no change of leucine-309.

View larger version (75K): [in this window] [in a new window]   Figure 2. DNA sequence analysis of the V2 receptor gene in a healthy man and in a patient with NDI. A point mutation (C to T) was detected at codon 104 in the patient. This missense mutation substitutes cysteine for arginine (R104C).

View larger version (39K): [in this window] [in a new window]   Figure 3. Model of the human vasopressin V2 receptor and location of the R104C mutation. The putative seven-transmembrane structure of the V2 receptor and the site of mutation are indicated. Also depicted are a potential glycosylation site at asparagine 22, a disulfide bond between cysteines 112 and 192 (), and palmitoylation sites at cysteines 341 and 342. An R104C mutation is located in the first extracellular loop (•).

To characterize the insensitivity to AVP of the mutant receptors, we studied the binding properties of the wild-type, R104C, and R104S V2 receptors. We used the R104S mutant to estimate the significance of the cysteine-104 residue. Expression vectors, which introduced these receptor genes, were transfected into COS-7 cells, and the properties of ligand binding were examined.

For the benefit of more precise functional comparison of the wild-type and mutant receptors, we determined receptor mRNA contents of each cell line. The quantitative RT-PCR analysis showed mRNA expressions of each cell lines were same level statistically (data not shown). Thus, we considered these transfection efficiencies equal to each other.

Saturation experiments were performed on the three transfected COS-7 cell lines, and specific bindings were calculated from total and nonspecific bindings (Fig. 4A). The results of Scatchard linear transformation of the specific binding data are shown in Fig. 4B. We found that the binding sites of these receptors were in a single class and that their dissociation constants (K_D) and total binding capacities (B_max) could be estimated from the data. The analysis gave us interesting results (Table 1). The binding capacity of the R104C mutant was reduced to 10% of the wild type, but binding affinity was strong rather than wild type. On the other hand, binding capacity of R104S mutant receptor was recovered up to 50% of the wild type.

View larger version (14K): [in this window] [in a new window]   Figure 4. Binding of [3H]-AVP to V2 receptors in the wild-type, R104C, and R104S COS-7 cells. A, Saturation binding experiments were performed with increasing concentrations of [3H]-AVP in the presence or absence of 10 µM unlabeled AVP. Specific binding of the wild-type (•), R104C (), and R104S (x) V2 receptors is shown. B, Scatchard linear transformation of the data is shown.

View larger version (24K): [in this window] [in a new window]   Figure 5. AVP-induced cAMP production mediated by wild-type and two mutant V2 receptors with intracellular COS-7 cells. cAMP accumulation in the COS-7 cells expressing wild-type (•), R104C (), and R104S (x) receptors is shown. AVP dose-response curves were obtained with analysis software and determined the maximum increase in cAMP levels (above basal) and EC50.

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

 
We report a novel V2 receptor point mutation (R104C) in a patient with congenital NDI. The peptide substitution caused by this mutation occurs in the first extracellular loop of the receptor, and this position seems to be very important for receptor function. Transient expression studies with COS-7 cells showed that the receptor-binding capacity and AVP-induced cAMP accumulation reduced in comparison with the wild type. It suggests that R104C receptor dysfunction cause the renal tubular resistance for AVP. The receptor-binding capacity and cAMP accumulation was recovered by substitution of serine for cysteine-104. It seems that the sulfhydryl group of cysteine-104 is a primary cause of receptor dysfunction.

Among the vasopressin receptor subfamily, two conserved regions are found in the first and second extracellular loops. These regions are spatially continuous domains stabilized by a conserved disulfide bond and are thought to be part of peptide-recognition sites. Several mutations have been detected in the first or second extracellular loop in patients with X-linked recessive NDI, and the insensitivity of these mutant receptors to AVP were proven by expression studies (6, 7, 11, 12, 13, 14, 15, 16, 17, 18). The arginine-104, in the first extracellular loop of the human V2 receptor, is conserved among the mammalian vasopressin receptor subfamily, with the exception of the rat V1a receptor (19). Therefore, a mutation at this position would be expected to result in a substantial change of the receptor function. In fact, transient expression studies of R104C mutant showed that 10% of binding capacity and 50% of cAMP accumulation by comparison with wild type. It seems reasonable to suppose that the renal tubular resistance of the patient is caused by the R104C mutation.

Most of the missense mutations found in the first or second extracellular loop of the V2 receptor substitute a cysteine for the original amino acid; R106C, R181C, G185C, R202C, and Y205C are previously cited examples of such conversions (10, 11, 12, 13). The human V2 receptor has a predicted disulfide bond between cysteine-112 and cysteine-192 in the first and second extracellular loops, and a similar bond is conserved in the vasopressin and oxytocin receptor subfamilies (5, 20). Disulfide bonds are demonstrated in other G protein-coupled seven-transmembrane-domain receptors. It is probable that they are important for maintaining the structural and functional integrity of the receptors. Receptor dysfunction is caused by mutations that introduce a cysteine near the disulfide bond, which suggests that the new cysteine have the potential to interfere with the bond, perhaps by forming a false disulfide bond.

To determine the significance of the cysteine in the R104C mutant, we used another mutant V2 receptor whose arginine-104 was exchanged for serine (R104S). The structure of serine is similar to cysteine, but serine has a hydroxyl group instead of the sulfhydryl group that forms the disulfide bond. Because of the minimum structural changes between the polypeptides, many researchers have used serine to estimate the effect of the sulfhydryl group of cysteine. Thus, the R104S mutant may be an appropriate model of a mutant that has no sulfhydryl group on residue 104. Because of the substitution of serine for cysteine, V2 receptor-binding capacity was increased five times, and binding affinities made little difference. AVP-induced cAMP accumulation was also increased 1.5 times. Because the substitution improved the dysfunction of the R104C receptor, it is clear that the sulfhydryl group of the cysteine-104 is a primary cause of the dysfunction.

Most mutations in which a cysteine replaces another amino acid in the first or second extracellular loop show decreased affinities of ligand binding (6, 7). Yet, our results showed that the binding affinity of R104C mutant was not decreased but had severely decreased binding capacity compared with the wild type. It suggests that the loss of binding capacity is caused by the reduction of active receptors on the cell surface. Similar results of expression studies have been obtained with V2 receptor mutants G201D, R202C, and C341S/C342S (21, 22, 23, 24). These experiments reached the conclusion that the transport of the mutant receptors on the cell surface was affected. We did not evaluate the transport of R104C mutant receptor by measuring the protein distribution in expressing cells. The future direction of our investigations will be to assess the receptor distribution in transfected cells.

In conclusion, we have detected a novel V2 receptor mutation and demonstrated that the dysfunction of the mutant receptor was caused by reduction of ligand-binding capacity and adenylyl cyclase activity. We have also demonstrated that the sulfhydryl group of the cysteine at residue 104 of the mutant receptor is primarily responsible for the dysfunction.

	Acknowledgments

 
We thank the patient for his participation and Dr. Shigeru Matsukawa for technical support. We also thank Drs. Kenzou Uchida, Yoshiyu Takeda, and Yuji Ito for their cooperation in our studies.

	Footnotes

 
Address correspondence and requests for reprints to: Satoru Inaba, M.D., Third Department of Internal Medicine, Fukui Medical University, 23-1 Shimoaitsuki, Matsuoka, Fukui 910-1193, Japan.

Received March 21, 2000.

Revised August 29, 2000.

Accepted October 3, 2000.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Kambouris M, Dlouhy SR, Trofatter JA, Conneally PM, Hodes ME. 1988 Localization of the gene for X-linked nephrogenic diabetes insipidus to Xq28. Am J Med Genet. 29:239–246.[CrossRef][Medline]
2. Knoers N, van der Heyden H, van der Oost BA, Monnens L, Willems J, Ropers HH. 1988 Nephrogenic diabetes insipidus: close linkage with markers from the distal long arm of the human X chromosome. Hum Genet. 30:31–38.
3. Birnbaumer M, Seibold A, Gilbert S, et al. 1992 Molecular cloning of the receptor human antidiuretic hormone. Nature. 357:333–335.[CrossRef][Medline]
4. Seibold A, Brabet P, Rosenthal W, Birnbaumer M. 1992 Structure and chromosomal localization of the human antidiuretic hormone receptor gene. Am J Hum Genet. 51:1078–1083.[Medline]
5. Lolait SJ, O’Carroll AM, McBride OW, Konig M, Morel A, Brownstein MJ. 1992 Cloning and characterization of a vasopressin V2 receptor and possible link to nephrogenic diabetes insipidus. Nature. 357:336–339.[CrossRef][Medline]
6. Pan Y, Wilson P, Gitschier J. 1994 The effect of eight V2 vasopressin receptor mutations on stimulation of adenylyl cyclase and binding to vasopressin. J Biol Chem. 269:31933–31937.[Abstract/Free Full Text]
7. Yokoyama K, Yamaguti A, Izumi M, et al. 1996 A low affinity vasopressin V2 receptor gene in a kindred with X-linked nephrogenic diabetes insipidus. J Am Soc Nephrol. 7:410–414.[Abstract]
8. Mouillac B, Chini B, Balestre MN, et al. 1995 The binding site of neuropeptide vasopressin V1a receptor. J Biol Chem. 43:25771–25777.
9. Luthman H, Magnusson G. 1983 High efficiency polyoma DNA transfection of chloroquine-treated cells. Nucleic Acids Res. 11:1295–1308.[Abstract/Free Full Text]
10. Pan Y, Metzenberg A, Das S, Jing B, Gitschier J. 1992 Mutations in the V2 vasopressin receptor gene are associated with X-linked nephrogenic diabetes insipidus. Nat Genet. 2:103–106.[CrossRef][Medline]
11. van den Ouweland AM, Dreesen JC, Verdijk M, et al. 1992 Mutation in the vasopressin type 2 receptor gene (AVPR2) associated with nephrogenic diabetes insipidus. Nat Genet. 2:99–102.[CrossRef][Medline]
12. Wilden RS, Antush MJ, Bennett RL, Schoof JM, Scott CR. 1994 Heterogeneous AVPR2 gene mutations in congenital nephrogenic diabetes insipidus. Am J Hum Genet. 55:266–77.[Medline]
13. Bichet DG, Birnbaumer M, Lonegan M, et al. 1994 Nature and recurrence of AVPR2 mutations in X-linked nephrogenic diabetes insipidus. Am J Hum Genet. 55:278–286.[Medline]
14. Bichet DG, Arthus MF, Lonegan M, et al. 1993 X-linked nephrogenic diabetes insipidus mutations in North America and the Hopewell hypothesis. J Clin Invest. 92:1262–1268.
15. Holtzman EJ, Harris HWH, Kolakowski LF, Guay-Woodford LM, Botelho B, Ausiello DA. 1993 A molecular defect in the vasopressin V2 receptor gene causing nephrogenic diabetes insipidus. N Engl J Med. 328:1534–1537.[Free Full Text]
16. Knoers NVAM, van den Ouweland AMW, Verdijk M, Monnens LAH, van Oost BA. 1994 Inheritance of mutations in the V2 receptor gene in thirteen families with nephrogenic diabetes insipidus. Kidney Int. 46:170–176.[Medline]
17. Birnbaumer M, Gilbert S, Rosenthal W. 1994 An extracellular congenital nephrogenic diabetes insipidus mutation of the vasopressin receptor reduces cell surface expression, affinity for ligand, and coupling to the Gs/adenylyl cyclase system. Mol Endocrinol. 8:886–894.[Abstract]
18. Tsukaguchi H, Matsubara H, Aritaki S, Kimura T, Abe S, Inada M. 1993 Two novel mutations in the vasopressin V2 receptor gene in unrelated Japanese kindreds with nephrogenic diabetes insipidus. Biochem Biophys Res Commun. 197:1000–1010.[CrossRef][Medline]
19. Morel A, O’Carroll AM, Brownstein MJ, Lolait SJ. 1992 Molecular cloning and expression of a rat V1a arginine vasopressin receptor. Nature. 356:523–526.[CrossRef][Medline]
20. Kimura T, Tanizawa O, Mori K, Brownstein MJ, Okayama H. 1992 Structure and expression of a human oxytocin receptor. Nature. 356:526–529.[CrossRef][Medline]
21. Tsukaguchi H, Matsubara H, Inada M. 1995 Expression studies of two vasopressin V2 receptor gene mutations R202C and 804insG in nephrogenic diabetes insipidus. Kidney Int. 48:554–562.[Medline]
22. Sadeghi H, Robertson GL, Bichet DG, Innamorati G, Birnbaumer M. 1997 Biochemical basis of partial nephrogenic diabetes insipidus phenotypes. Mol Endocrinol. 11:1806–1813.[Abstract/Free Full Text]
23. Schulein R, Liebenhoff U, Muller H, Birnbaumer M, Rosenthal W. 1996 Properties of the human arginine vasopressin V2 receptor after site-directed mutagenesis of its putative palmitoylation site. Biochem J. 313:611–616.
24. Tsukaguchi H, Matsubara H, Taketani S, Mori Y, Seido T, Inada M. 1995 Binding-, intracellular transport-, and biosynthesis-defective mutants of vasopressin type 2 receptor in patients with X-linked nephrogenic diabetes insipidus. J Clin Invest. 96:2043–2050.

This article has been cited by other articles:

				J. J. Bedford, S. Weggery, G. Ellis, F. J. McDonald, P. R. Joyce, J. P. Leader, and R. J. Walker Lithium-induced Nephrogenic Diabetes Insipidus: Renal Effects of Amiloride Clin. J. Am. Soc. Nephrol., September 1, 2008; 3(5): 1324 - 1331. [Abstract] [Full Text] [PDF]

				J. H. Robben, N. V. A. M. Knoers, and P. M. T. Deen Cell biological aspects of the vasopressin type-2 receptor and aquaporin 2 water channel in nephrogenic diabetes insipidus. Am J Physiol Renal Physiol, August 1, 2006; 291(2): F257 - F270. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Submit a related Letter to the Editor Purchase Article View Shopping Cart Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Inaba, S. Articles by Miyamori, I. Search for Related Content PubMed PubMed Citation Articles by Inaba, S. Articles by Miyamori, I.