Dual roles for glucokinase in glucose homeostasis as determined by liver and pancreatic beta cell-specific gene knock-outs using Cre recombinase.
Glucokinase (GK) gene mutations cause diabetes mellitus in both humans and mouse models, but the pathophysiological basis is only partially defined. We have used cre-loxP technology in combination with gene targeting to perform global, beta cell-, and hepatocyte-specific gene knock-outs of this enzyme in mice. Gene targeting was used to create a triple-loxed gk allele, which was converted by partial or total Cre-mediated recombination to a conditional allele lacking neomycin resistance, or to a null allele, respectively. beta cell- and hepatocyte-specific expression of Cre was achieved using transgenes that contain either insulin or albumin promoter/enhancer sequences. By intercrossing the transgenic mice that express Cre in a cell-specific manner with mice containing a conditional gk allele, we obtained animals with either a beta cell or hepatocyte-specific knock-out of GK. Animals either globally deficient in GK, or lacking GK just in beta cells, die within a few days of birth from severe diabetes. Mice that are heterozygous null for GK, either globally or just in the beta cell, survive but are moderately hyperglycemic. Mice that lack GK only in the liver are only mildly hyperglycemic but display pronounced defects in both glycogen synthesis and glucose turnover rates during a hyperglycemic clamp. Interestingly, hepatic GK knock-out mice also have impaired insulin secretion in response to glucose. These studies indicate that deficiencies in both beta cell and hepatic GK contribute to the hyperglycemia of MODY-2.
Postic
C
C
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
Shiota
M
M
Niswender
K D
KD
Jetton
T L
TL
Chen
Y
Y
Moates
J M
JM
Shelton
K D
KD
Lindner
J
J
Cherrington
A D
AD
Magnuson
M A
MA
D000418:Albumins / Q000235:genetics; D000483:Alleles; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D017353:Gene Deletion; D005941:Glucokinase / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D006706:Homeostasis; D007328:Insulin / Q000235:genetics; D019426:Integrases / Q000235:genetics*; D007515:Islets of Langerhans / Q000201:enzymology / Q000378:metabolism*; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D019076:Transgenes; D014764:Viral Proteins
D000418:Albumins; D017931:DNA Primers; D007328:Insulin; D014764:Viral Proteins; D005941:Glucokinase; C045073:Cre recombinase; D019426:Integrases; D005947:Glucose