A murine model of autosomal dominant neurohypophyseal diabetes insipidus reveals progressive loss of vasopressin-producing neurons
Theron A. Russell, … , Jeffrey Weiss, J. Larry Jameson
Theron A. Russell, … , Jeffrey Weiss, J. Larry Jameson
Published December 1, 2003
Citation Information: J Clin Invest. 2003;112(11):1697-1706. https://doi.org/10.1172/JCI18616.
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A murine model of autosomal dominant neurohypophyseal diabetes insipidus reveals progressive loss of vasopressin-producing neurons

Abstract

Familial neurohypophyseal diabetes insipidus (FNDI) is an autosomal dominant disorder caused by mutations in the arginine vasopressin (AVP) precursor. The pathogenesis of FNDI is proposed to involve mutant protein–induced loss of AVP-producing neurons. We established murine knock-in models of two different naturally occurring human mutations that cause FNDI. A mutation in the AVP signal sequence [A(–1)T] is associated with a relatively mild phenotype or delayed presentation in humans. This mutation caused no apparent phenotype in mice. In contrast, heterozygous mice expressing a mutation that truncates the AVP precursor (C67X) exhibited polyuria and polydipsia by 2 months of age and these features of DI progressively worsened with age. Studies of the paraventricular and supraoptic nuclei revealed induction of the chaperone protein BiP and progressive loss of AVP-producing neurons relative to oxytocin-producing neurons. In addition, Avp gene products were not detected in the neuronal projections, suggesting retention of WT and mutant AVP precursors within the cell bodies. In summary, this murine model of FNDI recapitulates many features of the human disorder and demonstrates that expression of the mutant AVP precursor leads to progressive neuronal cell loss.

Authors

Theron A. Russell, Masafumi Ito, Mika Ito, Richard N. Yu, Fred A. Martinson, Jeffrey Weiss, J. Larry Jameson

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Figure 1

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Targeted mutagenesis of the mouse Avp gene. (a) Targeting strategy. Spec...
Targeted mutagenesis of the mouse Avp gene. (a) Targeting strategy. Specific mutations and restriction sites were inserted into exon 1 [A(–1)T; ScaI] or exon 2 [C67X; NheI] by homologous recombination. An additional XbaI site (X*) is created after Cre excision of the loxP-Neo-loxP cassette from the A(–1)T targeted allele. Introduced restriction sites were used to detect mutant and WT Avp genes and reverse-transcribed Avp mRNA. White boxes, Avp gene exons; gray boxes, Oxt gene exons. X, XbaI; H, HindIII; E, EcoRI; A, AccI. (b) Southern blot analysis. XbaI- and ScaI-digested genomic DNA was hybridized with a 1,214-bp probe (HindIII-EcoRI DNA fragment), labeling a 2,375-bp XbaI-digested DNA for the normal allele and 1,507-bp (XbaI-ScaI) and 404-bp (ScaI-XbaI*) fragments for the A(–1)T mutant allele. Digestion with EcoRI and NheI and hybridization with a 912-bp probe (XbaI-AccI DNA fragment) labeled a 4,578-bp EcoRI-digested DNA for the normal allele and 978-bp (EcoRI-NheI) and 3,600-bp (NheI-EcoRI) DNAs for the C67X mutant allele. (c) RT-PCR analysis for the detection of WT and mutant Avp transcripts in the hypothalamus. A 366-bp cDNA spanning the A(–1)T mutation was amplified using forward and reverse primers located within exon 1 and exon 2, respectively. Restriction digestion with ScaI generated a 366-bp band from the normal allele and 265- and 101-bp fragments from the mutant allele. DNA spanning the C67X mutation (267 bp) was amplified by the use of forward (exon 2) and reverse (exon 3) primers. NheI digestion gave rise to 163- and 104-bp fragments derived from the mutant allele.