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The Journal of Clinical Endocrinology & Metabolism Vol. 83, No. 11 3958-3964
Copyright © 1998 by The Endocrine Society

Original Studies

Two Novel Mutations of the Vasopressin Gene Associated with Familial Diabetes Insipidus and Identification of an Asymptomatic Carrier Infant1

Frederick D. Grant, Arshanoush Ahmadi, Catherine M. Hosley and Joseph A. Majzoub

Endocrinology-Hypertension Division, Brigham and Women’s Hospital (F.D.G., C.M.H.), Division of Endocrinology and the Mental Retardation Research Center, The Children’s Hospital (F.D.G., J.A.M.), Program in Medical Science, Boston University School of Medicine (A.A.), and the Departments of Medicine (F.D.G., J.A.M.) and Pediatrics (J.A.M), Harvard Medical School, Boston, Massachusetts 02115

Address all correspondence and requests for reprints to: Frederick D. Grant, M.D., Endocrinology-Hypertension Division, Brigham and Women’s Hospital, 221 Longwood Avenue, Boston, Massachusetts 02115. E-mail: grantf{at}a1.tch.harvard.edu


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Familial diabetes insipidus (FDI) is a syndrome of central vasopressin deficiency that is inherited in an autosomal dominant manner and that typically becomes clinically apparent in the first decade of life. Two novel mutations of the vasopressin gene have been identified in two previously unstudied kindreds with FDI. In each kindred, the inheritance of the FDI phenotype was consistent with an autosomal dominant mode of inheritance. In each proband, the diagnosis of central diabetes insipidus had been confirmed previously with a water deprivation protocol. After extraction of genomic DNA from each individual, the three exons of the vasopressin gene were separately amplified by PCR and directly sequenced using an automated dye termination method. In the proband and two other carriers of one kindred, a heterozygous C to T mutation was identified at nucleotide 1857. This is predicted to produce a serine to phenylalanine substitution at residue 56 of the vasopressin-related neurophysin peptide encoded by the mutated allele. The mutation also abolished an MspI site in the vasopressin sequence, and analysis of genomic DNA from eight members of the kindred (five with FDI) confirmed segregation of the mutation with the FDI phenotype. Another member of the kindred, a 13-month-old infant, also has the heterozygous C to T mutation, but a formal water balance study showed no evidence of diabetes insipidus. In the proband of the other kindred, a heterozygous G to A mutation was identified at nucleotide 1873. This mutation would be predicted to cause a cysteine to tyrosine substitution at residue 61 of the neurophysin encoded by the mutated allele. This heterozygous mutation was confirmed by the presence of an RsaI restriction site in one vasopressin allele in two members of the kindred. Therefore, two novel heterozygous mutations of the vasopressin gene have been identified in FDI kindreds. In one kindred, an asymptomatic carrier infant was identified and will require continued observation to determine whether she will develop clinical diabetes insipidus. The presence of these two novel mutations in a region of the vasopressin gene where other FDI mutations have been reported suggests that the part of the neurophysin peptide encoded by these sequences may be critically important in the appropriate expression of vasopressin.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
FAMILIAL diabetes insipidus (FDI) is a syndrome of central vasopressin deficiency that is inherited as an autosomal dominant trait with a high degree of penetrance (1, 2). Affected individuals typically exhibit no evidence of diabetes insipidus in infancy and then develop clinical symptoms of vasopressin deficiency in the first decade of life. Clinical studies of individuals with FDI have confirmed a deficiency of circulating vasopressin (3, 4).

The vasopressin gene contains three exons that encode the vasopressin preprohormone (5). Exon A contains sequences encoding a signal peptide and the nine-amino acid vasopressin peptide. The vasopressin-related neurophysin is encoded by exon B together with small contiguous regions of exons A and C. Exon C also encodes the vasopressin-related glycopeptide. Since 1991, a number of mutations in the vasopressin gene have been reported to be associated with FDI (6). Nearly all of these mutations have been located in the neurophysin and signal peptide domains of the vasopressin gene, although there have been two preliminary reports of mutations in the region encoding the vasopressin nonapeptide (7, 8). No asymptomatic polymorphisms of the vasopressin gene have been found. Although clinical studies have demonstrated symptoms of diabetes insipidus in previously asymptomatic members of FDI kindreds, the identification of a mutation in an individual before the development of symptomatic diabetes insipidus may guide clinical follow-up and treatment and prevent the morbidity of untreated diabetes insipidus in an infant.

In the present study, we report two previously unstudied kindreds with FDI in which analysis of the vasopressin gene sequence has revealed two novel mutations in exon B. In one kindred, the proband appears to carry a de novo mutation. In the other kindred, individuals from multiple generations have been studied, and an asymptomatic infant has been identified as a carrier of a heterozygous mutation.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Clinical history

Two previously unstudied FDI kindreds (Fig. 1Go) were identified from patients referred to the endocrinology clinic at Children’s Hospital. Vasopressin levels had been measured by a RIA with a reported reference range of 1–13 pg/mL (Nichols Institute Diagnostics, Riveredge, NJ). The kindreds are not known to be related, and both are of northern European ancestry.



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  		Figure 1. Pedigrees of two kindreds with autosomal dominant familial diabetes insipidus. Affected individuals are indicated by shaded symbols. Males and females are indicated by squares and circles, respectively. Oblique squares represent multiple family members with no reported history of diabetes insipidus. In kindred A, genetic analysis was performed in the nine family members (A–J), and in kindred B, genetic analysis was performed in two individuals (A and B).

 
Kindred A. On the basis of a family history, the proband (A) was first suspected to have diabetes insipidus when he developed polyuria and polydipsia at age 14 months. On one occasion, basal serum osmolality was 279 mosmol/kg, and basal urine osmolality was 101 mosmol/kg. After an overnight fast and water deprivation, serum osmolality was 294 mosmol/kg, and urine osmolality was 312 mosmol/kg with a plasma vasopressin concentration of 1.4 pg/mL. This was interpreted as showing no evidence of overt vasopressin deficiency. However, based on the family history and the clinical presentation, the patient was begun on desmopressin therapy during the subsequent year. The patient responded with resolution of symptomatic polyuria, an increase in growth rate, and improvement of hydronephrosis. A 6-h water deprivation study off desmopressin therapy at age 4 yr showed a maximal serum osmolality of 306 mosmol/kg and maximal urine osmolality of 90 mosmol/kg with a plasma vasopressin level of less than 1.0 pg/mL. The proband’s mother (G) had been diagnosed with diabetes insipidus at age 6 yr, and the family history of familial diabetes insipidus extends over six generations. Nine individuals (A–J) in this kindred, including five with a clinical diagnosis of FDI, underwent genetic analysis.

Kindred B. The diagnosis of central diabetes insipidus was made in the proband (B) in 1958 at age 19 months after he presented with 8 months of polyuria and polydipsia of up to 2 qt/day. There was no previous family history of diabetes insipidus. After water deprivation, maximum urine specific gravity was 1.010. With hypertonic saline infusion, maximum urine osmolality was 85 mosmol/kg, which increased to 316 mosmol/kg after administration of posterior pituitary extract. The patient was treated with posterior pituitary extract (and more recently with desmopressin by rhinal tube) with correction of the symptoms of diabetes insipidus. The proband’s daughter developed polyuria and polydipsia at approximately 2 yr of age. By age 5 yr she was drinking up to 6 qt/day. Polyuria and polydipsia responded to treatment with intranasal desmopressin. Evaluation off desmopressin after 2.5 h of water deprivation showed a serum osmolality of 293 mosmol/kg, urine osmolality of 154 mosmol/kg, and plasma vasopressin of less than 1.0 pg/mL. The proband’s son developed polyuria and polydipsia of 60–70 oz/day at age 18 months. After 4 h of water deprivation with loss of 3% of body weight, urine specific gravity remained 1.001 with an osmolality of 150 mosmol/kg. A plasma vasopressin level was less than 1.0 pg/mL, with a plasma osmolality of 293 mosmol/kg. Both offspring responded to nasally administered desmopressin with an amelioration of the symptoms of diabetes insipidus. Genetic analysis of the proband and his son was performed, but other family members chose not to undergo genetic analysis.

DNA isolation

Genomic DNA was isolated from peripheral blood leukocytes following standard procedures. Peripheral venous blood was obtained by venipuncture into glass tubes containing sodium ethylenediamine tetraacetate (EDTA) and stored at 4 C before extraction. Seven milliliters of blood were mixed with an equal volume of nuclei extraction buffer [64 mmol/L sucrose (Life Technologies, Gaithersburg, MD), 2 mmol/L Tris at pH 7.6, 1 mmol/L magnesium chloride, and 2% Triton X-100 (all from Sigma Chemical Co., Inc., St. Louis, MO)] and incubated at 4 C for 3 h. Leukocyte nuclei were pelleted by centrifugation at 1000 x g for 15 min. The pellet was resuspended in 5 mL 24 mmol/L EDTA (Sigma) and 75 mmol/L NaCl (CMS Chempure Laboratories, Houston, TX) and homogenized through a 20-gauge needle. After addition of SDS (Bio-Rad, Hercules, CA) to a final concentration of 0.5% and proteinase K (American Bioanalytical, Natick, MA) to a final concentration of 0.5%, the DNA was incubated at 37 C for 17 h. The DNA was extracted progressively with phenol (Amresco, Solon, OH; equilibrated with 10 mmol/L Tris and 1 mmol/L EDTA at pH 7.6), a 25:1:24 mixture of phenol-isoamyl alcohol-chloroform, and finally chloroform (both from Fisher Scientific International, Inc., Fairlawn, NJ). The extracted DNA solution was dialyzed against four changes of a buffer containing 5 mmol/L EDTA, 10 mmol/L Tris (pH 7.6), and 10 mmol/L NaCl. The final DNA concentration was determined by spectrophotometric measurement of the OD260.

PCR

PCR amplification (9) of the three exons of the vasopressin gene was performed with oligonucleotide primers (Table 1Go) corresponding to intron sequences flanking each of the three exons (5). Each 100-µL amplification reaction (10) contained 100 ng genomic DNA; 30 pmol each of the appropriate forward (sense) and reverse (antisense) primers; 20 pmol each of deoxy-ATP (dATP), dCTP, dGTP, and dTTP; and 20 µL PCR buffer (buffer A; Invitrogen, San Diego, CA; 300 mmol/L Tris-HCl, 75 mmol/L ammonium sulfate, and 7.5 mmol/L magnesium chloride, pH 8.5). After 5 min of initial denaturation at 100 C, the reaction was cooled to 98 C, and 1 U Taq DNA polymerase (Boehringer Mannheim, Indianapolis, IN) was added. Thirty-five cycles of amplification were performed, with denaturation for 1 min at 96 C and annealing and extension for 5 min at 70 C, followed by one final extension for 5 min at 72 C.


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  		Table 1. Oligonucleotide primers used to PCR amplify vasopressin exons

 
DNA sequencing

In preparation for sequencing, PCR products were purified using a glass bead method (Wizard PCR Preps, Promega Corp., Madison, WI) and were directly sequenced using a Taq polymerase dideoxy termination method (Perkin-Elmer/Cetus, Norwalk, CT) and an automated sequencer (ABI/Perkin Elmer 373 DNA Sequencer) (11). Both strands of each PCR product were sequenced using the same upstream or downstream oligonucleotide primers that had been used for PCR amplification of each DNA fragment.

Restriction digestion analysis

PCR fragments were subjected to digestion with restriction enzymes chosen to identify polymorphisms in the mutated vasopressin sequence (12). Digestion products were analyzed by agarose gel electrophoresis using an ethidium-stained agarose gel made of 1% agarose (type I-A, Sigma) and 1% Nu-Sieve (FMC BioProducts, Rockland ME) in 90 mmol/L Tris base, 90 mmol/L boric acid (Sigma), and 2 mmol/L EDTA. The gel was visualized under UV light and recorded on Polaroid 57 Instant Sheet Film (ASA 3000, Madison, WI).

Water balance studies

The individual identified as a heterozygous carrier of a vasopressin gene mutation underwent an outpatient study of water balance (13). The individual was fasted with no fluid intake after midnight and arrived for formal study at 0800 h. After placement of an in-dwelling iv catheter, baseline serum sodium level and osmolality and urine osmolality were determined. Weights were assessed hourly, serum sodium and osmolality were assayed every hour, and urine samples for determination of osmolality were obtained with each void, with urine collected in an externally applied bag. Plasma for vasopressin was obtained at the beginning and conclusion of the study. Fluid restriction was to be continued until there was an obvious diagnosis or a loss of more than 3% of basal weight.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Each of the three exons of the AVP gene was amplified and sequenced using DNA samples obtained from three affected and one nonaffected member of kindred A and from the proband of kindred B. Direct sequencing of the PCR-amplified exons revealed no differences in exon A or C compared with the published sequence of the human vasopressin gene (5). However, in the vasopressin gene of the affected individuals in each kindred, a unique heterozygous mutation was identified in the sequence of exon B.

In kindred A, direct sequencing of exon B of affected individuals A (the proband), G, and H identified a heterozygous C to T mutation at nucleotide 1857 (5) (Fig. 2AGo). This mutation predicts a serine (TCC) to phenylalanine (TTC) substitution at residue 56 of neurophysin. No polymorphisms were identified in the vasopressin sequence of the unaffected individual J.



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  		Figure 2. Direct sequencing of exon B PCR products obtained from the probands of each kindred compared to those from an unrelated normal control. A, In kindred A, a heterozygous C to T mutation was found at nucleotide 1857 of exon B of the vasopressin gene. Sequencing of the antisense strand showed heterozygous a G to A mutation complementary to the mutation observed in the sense strand PCR products. This mutation would predict a serine (ser) to phenylalanine (phe) substitution in the peptide encoded by the mutated allele. B, In the proband of kindred B, a G to A point mutation was identified at nucleotide 1873 of exon B. In the antisense strand, there was a complementary heterozygous C to T mutation. This mutation should result in replacement of the cysteine (cys) residue with a tyrosine (tyr) in the peptide encoded by the mutated allele.

 
The C to T mutation in affected individuals of kindred A should eliminate a MspI restriction site (12) on the affected allele. To confirm the segregation of the mutation with the FDI phenotype, PCR products obtained by amplification of exon B in nine members of the kindred were analyzed by MspI digestion and compared to known noncarrier DNA (Fig. 3Go). Digestion of PCR products obtained from the three individuals with heterozygous mutations of the vasopressin gene identified by sequence analysis showed a pattern consistent with the presence of both a mutated and a nonmutated allele. Analysis of DNA from five other kindred members confirmed segregation of the mutation with the FDI phenotype. The proband (A) and individuals D, F, G, and H have diabetes insipidus, and each has an MspI polymorphism consistent with a heterozygous mutation at nucleotide 1857 of the vasopressin gene. Individuals C, E, and J have no symptomatic abnormality of water balance and no evidence of a vasopressin gene mutation.



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  		Figure 3. Restriction fragment length polymorphism in the vasopressin gene resulting from a heterozygous mutation at nucleotide 1859 in kindred A. A, The vasopressin gene mutation identified in kindred A should abolish an MspI restriction site within the PCR fragment obtained from PCR amplification of exon B. Therefore, restriction digestion of the mutant allele should yield DNA fragments of 206 and 56 bp. MspI restriction digestion of the normal allele yields fragments of 83, 123, and 56 bp. B, After digestion with MspI, PCR products obtained from nine members of kindred A were analyzed by gel electrophoresis. The restriction pattern was consistent with the presence of the heterozygous mutation in samples from individuals A, B, D, F, G, and H. pl, Human vasopressin gene cloned into the pBR plasmid vector; nc, DNA from an unrelated normal control individual; mw, mol wt marker.

 
One additional member of kindred A without the clinical diagnosis of FDI also was found to carry the heterozygous mutation at nucleotide 1857 of the vasopressin gene. At the time of identification, this individual (B) was a 13-month-old female in whom the family had not noted any symptoms of polyuria or polydipsia. A formal water deprivation study was performed to assess this asymptomatic carrier infant for subclinical defects in water balance (13). At baseline, after an overnight fast, serum osmolality was 283 mosmol/kg, and serum sodium was 138 mEq/L. During an additional 4 h of water deprivation, maximal serum osmolality and sodium were 283 mosmol/kg and 140 mEq/L, respectively, and the peak plasma vasopressin level was 1.7 pg/mL. Urine osmolality determined approximately 2 h after initiation of the formal study was 713 mosmol/kg. A urine osmolality greater than 700 mosmol/kg and a plasma vasopressin level of 1.7 pg/mL at a plasma sodium level of 140 mEq/L demonstrated an appropriate antidiuretic response (14). At age 19 months, plasma sodium was 140 mEq/L, and urine specific gravity was 1.007 after overnight water deprivation.

In the proband (individual B) of kindred B, direct sequencing of exon B identified a heterozygous G to A mutation at nucleotide 1873 (5) (Fig. 2BGo). This mutation predicts a cysteine (TGC) to tyrosine (TAC) mutation at residue 61 of the neurophysin peptide. This mutation produces a new RsaI restriction site (12) in the affected allele. Digestion with RsaI of the PCR amplification product from exon B showed both digested and undigested fragments, consistent with a heterozygous mutation (Fig. 4Go). A similar pattern consistent with a heterozygous mutation at nucleotide 1873 was observed after RsaI digestion of the PCR product from exon B of individual A of this kindred. Other members of the kindred did not undergo genetic analysis.



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  		Figure 4. Restriction fragment length polymorphism in the vasopressin gene resulting from a heterozygous mutation at nucleotide 1873 in kindred B. A, The vasopressin gene mutation identified in kindred B is predicted to produce a new RsaI restriction site within the 284 bp PCR fragment obtained by amplifying exon B. Therefore, restriction digestion of the mutant allele should yield DNA fragments of 206 and 56 bp, whereas no digestion of the normal allele should occur. B, Gel electrophoresis of RsaI-digested PCR products revealed a 217-bp fragment as well as an uncut 284-bp fragment in samples from individuals A and B, which were consistent with a heterozygous mutation in the vasopressin gene. Only the uncut 284-bp fragment was produced in PCR products amplified from normal control genomic DNA (NC).

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Since 1991, a number of mutations of the vasopressin gene have been reported to be associated with FDI. Ito et al. (15) reported a heterozygous G to A mutation at nucleotide 1859 of exon B predictive of a glycine to serine mutation at residue 57 of neurophysin. Soon after, in a study of a Dutch kindred, Bahnsen found a G to T substitution at nucleotide 1740 of the vasopressin gene (16). This mutation also predicted a substitution of glycine with valine at residue 17 of the vasopressin-related neurophysin. Published studies have reported 22 different mutations associated with symptomatic FDI in 28 different kindreds (6, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25). Three mutations have been found in more than one presumed independent kindred (6).

The current study reports two additional novel mutations associated with FDI. In both kindreds, the diagnosis of diabetes insipidus was made by provocative studies in one or more individuals. In one kindred, the autosomal dominant inheritance pattern is evident with the identification of individuals with symptomatic FDI in a six-generation pedigree. In the other kindred, diabetes insipidus may have resulted from a de novo mutation in the vasopressin gene of the proband, as there is no history of diabetes insipidus in the family before his presentation.

This study identified an asymptomatic infant that carries a copy of the mutant allele associated with FDI in other members of her kindred. This infant has no clinical symptoms of FDI and has demonstrated no abnormality of water balance during a formal water deprivation study and subsequent follow-up. Although it is possible that this individual will remain an asymptomatic carrier, her family history suggests that it is most likely that she will develop diabetes insipidus as she grows older. Previous clinical reports of FDI have suggested that the age of onset of symptoms of FDI is not consistent among individuals of the same kindred (1, 4, 26, 27). One previous report described development of diabetes insipidus in a previously asymptomatic infant who was later shown to have a heterozygous mutation in the vasopressin gene (19). The variable ages of onset may represent biological variability of the mutation or may reflect chronic variations in exposure to stimuli of vasopressin secretion. Knowledge of this infant’s carrier state should allow her family and physicians to identify the onset of symptoms sooner in the course of FDI. Prior genetic diagnosis may help prevent any potential morbidity associated with untreated diabetes insipidus in a child.

In both kindreds, the newly identified mutations alter the sequence of the vasopressin-related neurophysin (5). Similarly, all previously published reports of FDI mutations have found mutations in the sequences that encode either the neurophysin (6, 15, 16, 20, 22, 23, 25) or the signal peptide (6, 17, 18, 19, 21, 24) domains of the vasopressin preprohormone. Two recent preliminary reports have described mutations within the sequence encoding the nine-amino acid vasopressin peptide (7, 8). Interestingly, one of these mutations (which substitutes leucine for proline 7 of vasopressin) results in a recessive inheritance pattern of vasopressin deficiency (8). This distribution of mutated loci suggests that both the signal peptide and neurophysin are important for the appropriate expression and secretion of vasopressin.

Vasopressin and neurophysin are synthesized as part of a common prohormone and are secreted from the posterior pituitary in equimolar amounts (28, 29). The two mutations identified in the present report are located in regions of the neurophysin sequence in which other mutations have been identified. Three different nucleotide substitutions have been identified at position 1859 (6, 15) in close proximity to the C to T substitution at position 1857 in kindred A. As in kindred B, two other mutations previously have been reported to alter the cysteine at residue 61 (6). The heterogeneous distribution of mutations within the neurophysin sequence may suggest that the regions with symptomatic mutations have been identified because these regions are critical to the function of neurophysin. Polymorphisms occurring in regions less important to vasopressin secretion may remain undetected. However, it is also possible that the regions with identified mutations may be mutational "hot spots," with an increased susceptibility to mutation (30, 31).

Magnetic resonance and crystallography studies have suggested that the neurophysin molecule contains a binding site for vasopressin (32, 33). One of the vasopressin mutations associated with FDI is a trinucleotide deletion in the putative vasopressin-binding region of the neurophysin sequence (20). Although this is consistent with an alteration of vasopressin binding as one cause of FDI, it does not address the mechanism by which vasopressin mutations act in a dominant manner to cause vasopressin deficiency.

The large number of different mutations identified in association with FDI argues against a specific gain of function effect impairing vasopressin secretion. As some evidence suggests that neurophysin may form either dimers or tetramers (34, 35), a variety of mutations that disrupt the neurophysin structure could cause a transdominant negative effect by inhibiting appropriate multimer formation (36, 37). If multimer formation is important to vasopressin secretion, then disruption due to this transdominant effect could result in vasopressin deficiency. A variety of mutations could result in misfolding of the vasopressin prohormone. For example, in affected individuals of kindred B, the replacement of a cysteine with tyrosine could greatly alter the folding of the vasopressin prohormone. This could cause alteration in the processing or intracellular transport of vasopressin and lead to impaired secretion of appropriately processed vasopressin (36). However, this mechanism alone does not explain the asymptomatic nature of the disease early in life.

In vitro studies in cultured AtT-20 (38) and neuro2a (36, 39) cells have suggested that there is impairment of vasopressin secretion in cells expressing mutated vasopressin constructs. However, in the absence of coexpression of normal and mutant vasopressin constructs, the dominant nature of this mechanism could not be confirmed. In neuro2a cells differentiated by application of valproic acid, expression of mutated vasopressin constructs increased cell death (36, 39). Although this is supportive of the hypothesis that cell death occurs in vivo as a result of expression of a mutant vasopressin, these experiments have not demonstrated that impaired vasopressin secretion is dependent on prior neuronal cell death.

A small number of postmortem histological studies of the hypothalamus in individuals with FDI (40, 41, 42) have suggested that FDI involved a selective loss of vasopressin-expressing nerve cells in the hypothalamus. In contrast, Forssman reported finding no hypothalamic abnormalities on postmortem exam of an individual with inherited diabetes insipidus (26). Bergeron et al. (43) noted the loss of magnocellular vasopressin-expressing cells, but observed the presence of smaller vasopressin-expressing neurons. In another autopsy study by Nagai (44), the hypothalamic architecture was reported to be normal, with a reported history of FDI. Decreased vasopressin immunostaining was seen in the paraventricular nuclei, but normal levels of vasopressin were detected in the supraoptic nuclei of the hypothalamus. Because none of these autopsy studies was performed in individuals with an identified mutation of the vasopressin gene, it is not possible to rigorously correlate histological findings with autosomal dominant FDI. Although these reports together with the in vitro studies in cultured cells are consistent with the hypothesis of neuronal degeneration as a cause of FDI (36, 37), they do not provide a definitive answer as to the role of neurodegeneration in the development of FDI.

In summary, two novel heterozygous mutations in the vasopressin gene have been identified in two kindreds with FDI. Pedigree analysis is consistent with an autosomal dominant inheritance of each. The location of these mutations in a region of the neurophysin sequence where other FDI mutations have been identified suggests that this region of the neurophysin molecule may be important in the appropriate secretion of vasopressin. However, the heterogeneous distribution of FDI mutations also could be consistent with mutational hot spots in the vasopressin gene sequence. In one kindred, an asymptomatic infant has been identified as a carrier of one of the mutations. Provocative testing of this individual at 13 months of age and follow-up at 19 months have not identified a defect in water balance or in the vasopressin response to water deprivation. Although it is possible that this individual will remain an asymptomatic carrier, continued observation as she grows older is likely to reveal development of symptomatic FDI. If so, then this would demonstrate the preclinical diagnosis of FDI by genetic screening. Diagnosis of FDI by genetic methods before the development of symptoms should aid in the early identification and treatment of FDI in infants.


   Acknowledgments
 
We thank Gregory Goodwin, M.D., for the initial clinical analysis of kindred B, and Ingrid Holm, M.D., for providing the DNA extraction protocol.


   Footnotes
 
1 This work was supported by NIH Grants DK-02159 (to F.D.G.) and HD-18655 (to the Mental Retardation Research Center, The Children’s Hospital), a Basic Research Grant from the March of Dimes Birth Defects Foundation (to F.D.G.), and the Program in Medical Sciences of the Boston University School of Medicine (to A.A.). Presented in part at the 79th Annual Meeting of The Endocrine Society, Minneapolis, MN, June, 1997. Back

Received March 9, 1998.

Revised August 5, 1998.

Accepted August 11, 1998.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

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