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J. Biol. Chem., Vol. 265, Issue 36, 22371-22379, 12, 1990
MM McGrane, JS Yun, AF Moorman, WH Lamers, GK Hendrick, BM Arafah, EA Park, TE Wagner and RW Hanson
Transgenic mice were used to investigate sequences within the promoter of
the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP)
from the rat (EC 4.1.1.32) (PEPCK) which are involved in tissue- specific
and developmental regulation of gene expression. Segments of the PEPCK
promoter between -2000 and -109 were linked to the structural gene for
bovine growth hormone (bGH) and introduced into the germ line of mice by
microinjection. Bovine growth hormone mRNA was found in tissues that
express the endogenous PEPCK gene, mainly in the liver but to a lesser
extent in the kidney, adipose tissue, small intestine, and mammary gland.
In the liver the chimeric PEPCK/bGH(460) gene was expressed in periportal
cells, which is consistent with the zonation of endogenous PEPCK. The
PEPCK/bGH gene was not transcribed in the livers of fetal mice until
immediately before birth; at birth the concentration of bGH mRNA increased
200-fold. Our results indicate that the region of the PEPCK promoter from
-460 to +73 base pairs contains regulatory sequences required for
tissue-specific and developmental regulation of PEPCK gene expression. Mice
transgenic for PEPCK/bGH(460) were not hyperglycemic or hyperinsulinemic in
response to elevated bGH, as were transgenic mice with the MT/bGH gene. The
number of insulin receptors in skeletal muscle was no different in mice
transgenic for MT/bGH when compared with mice transgenic for PEPCK/bGH(460)
and control animals. However, mRNA abundance for the insulin-sensitive
glucose transporter in skeletal muscle was decreased in mice transgenic for
the MT/bGH gene. The differences in glucose homeostasis noted with the two
types of transgenic mice may be the result of the relative site of
expression, the different developmental pattern, or hormonal regulation of
expression of the bGH gene.
Metabolic effects of developmental, tissue-, and cell-specific expression of a chimeric phosphoenolpyruvate carboxykinase (GTP)/bovine growth hormone gene in transgenic mice
Pew Center for Molecular Nutrition, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106.
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