B6;129S6-Fam3b^tm1Bkht/J

Stock number: 013579
Research areas: Diabetes and Obesity Research

Description

Male mice homozygous for this knockout are glucose intolerant, have decreased hepatic insulin clearance and reduced hepatic glucose production in response to hyperinsulinemic conditions.

Detailed description

Homozygotes are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. No gene product (mRNA or protein) is detected by RT-PCR and Western blot analysis of pancreatic islets and small intestines from homozygotes. Male mice that are homozygous for the targeted mutation and 4 to 6 months of age exhibit higher blood glucose levels in glucose tolerance tests when compared to wildtype controls. Although, insulin sensitivity is normal in mutant mice, insulin levels are elevated after glucose stimulation (intraperitoneal glucose injection) due to decreased hepatic insulin clearance. Glucose intolerance is not observed in mutant female mice. Isolated pancreatic islets from male homozygotes have impaired glucose-stimulated insulin secretion and calcium signaling. Male homozygotes have reduced hepatic glucose production (HGP) and increased insulin-mediated HGP suppression conditions as determined by hyperinsulinemic-euglycemic clamp assays. Furthermore, male homozygotes display reduced expression of the critical gluconeogenic enzymes of glucose-6-phosphatase (G6P) and phosphoenolpyruvate carboxykinase (PEPCK) following a 24 hour fast as compared to wildtype controls as determined by RT-PCR.

Development

A targeting vector containing a Neo selection cassette was used to disrupt the transcriptional start site (TSS), secretion signal peptide (SP), and exons 1 and 2. The construct was electroporated into 129S6/SvEvTac derived TL1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were crossed to C57BL/6 mice. Heterozygotes were intercrossed to produce homozygotes. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.

Allele

Symbol: Fam3b^tm1Bkht
Name: targeted mutation 1, Brant Burkhardt
MGI accession ID: MGI:4882127
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Fam3b
Marker name: family with sequence similarity 3, member B
Chromosome: 16
Strain of origin: 129S6/SvEvTac
Molecular note: Exons 1 and 2 were replaced with a neomycin selection cassette in reverse orientation. This removed the transcriptional and translational start sites and the first 54 amino acids including the secretion signal peptide and helix A. mRNA was not detected in pancreatic islets, small intestine, liver, spleen, kidney, and brain by RT-PCR. Western blot analysis of pancratic islets demonstrated the absence of full length protein.