Hepatic lobular patterns of phosphoenolpyruvate carboxykinase, glycogen synthase, and glycogen phosphorylase in fasted and fed rats.
The goal of this study was to localize phosphoenolpyruvate carboxykinase (PEPCK), glycogen synthase (GS), and glycogen phosphorylase (GP) in the liver lobule by immunocytochemical techniques and to describe the effects of feeding and fasting on the distribution and quantity of these enzymes. Livers from ad lib fed and overnight fasted normal adult male rats were frozen in liquid nitrogen after transcardial perfusion with 30% sucrose. Serial cryostat sections of tissue were collected on slides, fixed by immersion in 4% paraformaldehyde, and incubated with antibodies against PEPCK, GS, and GP. Antibodies to these enzymes were visualized with a gold-conjugated secondary antibody and a silver enhancement technique. Fed animals demonstrated a periportal to pericentral gradient of PEPCK. Fasting increased the periportal content of PEPCK, induced the midlobular and centrilobular cells to express the enzyme, and steepened the periportal to pericentral gradient. The increase of PEPCK was confirmed by Western blot analysis. GS and GP were distributed throughout the lobule in the fed animal but often showed a centrilobular pattern, and fasting did not alter the lobular distribution of either enzyme. Western blot analysis revealed no changes in the amount of these enzymes in the fed or fasted state. The cellular distribution of the three enzymes is similar to that of hepatic glycogen, in that the immunoreactive material has a clumped appearance in the periportal hepatocytes and is more dispersed in the pericentral cells. On fasting the periportal hepatocytes lose the dense compact localization of the enzymes and the protein becomes more homogeneously distributed throughout the cytosol. Further studies are needed to elucidate the functional significance of the regional heterogeneity of the glycogen-metabolizing enzymes and the molecular mechanisms regulating their gene expression.
Giffin
B F
BF
Department of Anatomy and Cell Biology, University of Cincinnati College of Medicine, Ohio 45267-0521.
Drake
R L
RL
Morris
R E
RE
Cardell
R R
RR
D000818:Animals; D015153:Blotting, Western; D004435:Eating / Q000502:physiology*; D005215:Fasting / Q000502:physiology*; D006006:Glycogen Synthase / Q000032:analysis*; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology / Q000201:enzymology* / Q000502:physiology; D008112:Liver Glycogen / Q000032:analysis; D008297:Male; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000032:analysis*; D006005:Phosphorylases / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley
D008112:Liver Glycogen; D006005:Phosphorylases; D006006:Glycogen Synthase; D010729:Phosphoenolpyruvate Carboxykinase (GTP)