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. 1999 Aug 31;96(18):10152-7.
doi: 10.1073/pnas.96.18.10152.

Impaired glucose homeostasis and neonatal mortality in hepatocyte nuclear factor 3alpha-deficient mice

D Q Shih  1 , M A Navas, S Kuwajima, S A Duncan, M Stoffel
Affiliations

Impaired glucose homeostasis and neonatal mortality in hepatocyte nuclear factor 3alpha-deficient mice

D Q Shih et al. Proc Natl Acad Sci U S A. .

Abstract

Hepatocyte nuclear factors 3 (HNF-3) belong to an evolutionarily conserved family of transcription factors that are critical for diverse biological processes such as development, differentiation, and metabolism. To study the physiological role of HNF-3alpha, we generated mice that lack HNF-3alpha by homologous recombination in embryonic stem cells. Mice homozygous for a null mutation in the HNF-3alpha gene develop a complex phenotype that is characterized by abnormal feeding behavior, progressive starvation, persistent hypoglycemia, hypotriglyceridemia, wasting, and neonatal mortality between days 2 and 14. Hypoglycemia in HNF-3alpha-null mice leads to physiological counter-regulatory responses in glucocorticoid and growth hormone production and an inhibition of insulin secretion but fails to stimulate glucagon secretion. Glucagon-producing pancreatic alpha cells develop normally in HNF-3alpha-/- mice, but proglucagon mRNA levels are reduced 50%. Furthermore, the transcriptional levels of neuropeptide Y are also significantly reduced shortly after birth, implying a direct role of HNF-3alpha in the expression of these genes. In contrast, mRNA levels were increased in HNF-3 target genes phosphofructo-2-kinase/fructose-2,6-bisphophatase, insulin growth factor binding protein-1, and hexokinase I of HNF-3alpha-null mice. Mice lacking one or both HNF-3alpha alleles also show impaired insulin secretion and glucose intolerance after an intraperitoneal glucose challenge, indicating that pancreatic beta-cell function is also compromised. Our results indicate that HNF-3alpha plays a critical role in the regulation of glucose homeostasis and in pancreatic islet function.

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Figures

Figure 1
Figure 1
Disruption of the HNF-3α gene by homologous recombination. (A) Strategy for targeted disruption of the murine HNF-3α gene in ES cells. The genomic structure and restriction map of the mouse HNF-3α gene locus and targeting vector pPNT-2 used to disrupt the HNF-3α gene are shown. The targeting vector was constructed by deleting exon 2, including the DNA-binding domain downstream from the AflIII site and fusing it in-frame to the E. coli lacZ gene. The probe 3′ of the left targeting arm was used for Southern blot analysis and is shown as a bar. (B) Southern blot analysis of transfected ES cells and HNF3α+/− ES cells grown in the presence of G418. Genomic DNA was digested with HindIII (H3) and EcoRV. The wild-type allele shows a 7.5-kilobase band, and the targeted allele shows a 5.0-kilobase band.
Figure 2
Figure 2
Phenotype of HNF-3α mutant mice. (A) Growth curves of littermate wild-type +/+, heterozygous +/−, and homozygous HNF-3α−/− mice. (B) Stomach and duodenum of 2-day-old wild-type (+/+) and HNF-3α-null (−/−) mice. (C) Serum glucose concentrations in newborn (0), 3-, and 5-day-old mice. (D) Serum triglyceride and cholesterol levels. (E) Serum glucagon and insulin levels of 3-day-old mice. (F) Serum cortisol and growth hormone concentrations of 3- to 5-day-old littermates. (G) Measurements of serum glucose and insulin levels in 3-day-old mice 60 minutes after intraabdominal glucose administration. Blood glucose (H)and serum insulin (I)levels during an intraabdominal glucose administration in adult wild-type and heterozygous littermates. Numbers in brackets indicate number of animals studied. ∗, P < 0.05; ∗∗, P < 0.005; ∗∗∗, P < 0.0005.
Figure 3
Figure 3
Normal pancreatic morphology and immunostaining for glucagon and insulin in HNF-3α mice. Pancreatic tissue sections showing representative islets from 3-day-old pancreata of wild-type and HNF-3α mutant mice. Sections were stained with antibodies against glucagon and insulin as described in Materials and Methods. Glucagon and insulin are detected in the characteristic cell-lineage patterns. Magnification, ×400.
Figure 4
Figure 4
Steady-state mRNA levels of HNF-3 target genes. Steady-state mRNAs were measured by reverse transcription–PCR by using [α-32P]dCTP (16). PCR products were separated by PAGE, and bands were visualized by autoradiography. (A) Steady-state mRNA levles of genes that are reduced shortly after birth. (B) Steady state mRNA levels of hexokinase I, bifunctional enzyme phosphofructo-2-kinase/fructose-2,6-bisphosphatase-2 (PFK-2/FBase-2), and insulin growth factor binding protein-1 (IGFBP-1) are up-regulated in HNF-3α-null mice.
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References

    1. Lai E, Clark K L, Burley S K, Darnell J E., Jr Proc Natl Acad Sci USA. 1993;90:10421–10426. - PMC - PubMed
    1. McPherson C E, Shim E Y, Friedman D S, Zaret K S. Cell. 1993;75:387–398. - PubMed
    1. Shim E J, Woodcock C, Zaret K S. Genes Dev. 1998;12:5–10. - PMC - PubMed
    1. Costa R H, Grayson D R, Darnell J E., Jr Mol Cell Biol. 1989;9:1415–1426. - PMC - PubMed
    1. Lai E, Prezioso V R, Smith E, Litvin O, Costa R H, Darnell J E., Jr Genes Dev. 1990;4:1427–1436. - PubMed

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