Antizyme mRNA distribution and regulation in rat small intestinal enterocytes.
The protein ornithine decarboxylase antizyme (AZ) is inhibitory to both polyamine transport and synthesis. Experiments were performed to examine the distribution and regulation of AZ mRNA in cells of the small intestinal epithelium, a tissue exposed to high concentrations of extracellular polyamines and high levels of ornithine decarboxylase (ODC) activity. AZ mRNA was expressed in acutely isolated epithelial cells of rat jejunum and ileum; expression was higher in proximal than distal small intestine. In cells isolated from jejunal crypt-villus axis, AZ was expressed to high levels in cells from the small intestinal crypts but the message fell to near undetectable levels in cells of the villus tip. Western blot analysis demonstrated that distribution of AZ protein followed the distribution of AZ message. The distribution of ornithine decarboxylase activity along the crypt-villus axis was also determined. ODC activity and ODC protein were higher in cells from the upper villus than in cells isolated from the crypt. The intestinal lumen contains extremely high concentrations of free polyamines. The effect of depletion of endogenous polyamines or the addition of exogenous polyamines on AZ mRNA was evaluated in IEC-6 cells. Cells were depleted of intracellular polyamines by 72 hr of incubation in difluoromethylornithine. The fall in intracellular polyamine content was accompanied by a corresponding fivefold fall in AZ mRNA. When polyamine-depleted cells were treated with putrescine, the level of the AZ mRNA transcript was increased ninefold. These data demonstrate the expression of AZ gene in the longitudinal and crypt-villus axes of rat small intestine and show that AZ gene transcription is modulated by polyamines, an effect which may be involved in product suppression of polyamine synthesis.
Department of Physiology, School of Medicine, East Carolina University, Greenville, North Carolina 27858, USA.
J E
JE
Gill
J F
JF
Christian
E R
ER
Seidel
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003092:Colitis / Q000503:physiopathology*; D020895:Enterocytes / Q000378:metabolism*; D007082:Ileum / Q000166:cytology*; D007583:Jejunum / Q000166:cytology*; D008297:Male; D065108:Ornithine Decarboxylase Inhibitors; D011073:Polyamines / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014158:Transcription, Genetic
D065108:Ornithine Decarboxylase Inhibitors; D011073:Polyamines; D011506:Proteins; D012333:RNA, Messenger; C031017:ornithine decarboxylase antizyme