Differential effects of streptozotocin-induced diabetes on phenylalanine hydroxylase protein and mRNA abundance in isolated rat liver cells.
Phenylalanine hydroxylase catalyzes the major regulatory step of the phenylalanine degradation pathway. In view of the glucogenic nature of phenylalanine breakdown, and hence its potential contribution to glucose homeostasis, we have investigated the impact of streptozotocin-induced diabetes upon the expression of rat phenylalanine hydroxylase. Northern blot analysis revealed that induction of diabetes was associated with an increase in the in vivo abundance of hepatic phenylalanine hydroxylase-specific mRNA. This increase in mRNA abundance was maintained for at least 8 hr in liver cells isolated from diabetic animals. In contrast, phenylalanine hydroxylase immunoreactivity and enzymic activity decreased, over the 8 hr incubation period, to levels similar to those observed in liver cells from normal animals. These changes were retarded, but not prevented, by the presence of dexamethasone in incubation media. In liver cells from normal animals the abundance of phenylalanine hydroxylase-specific mRNA, immunoreactivity and enzymic activity, were largely insensitive to treatment with dexamethasone and/or glucagon over an 8 hr incubation period. It is concluded that, whereas diabetes-related alterations in phenylalanine hydroxylase-specific mRNA abundance persist after isolation of liver cells, changes in phenylalanine hydroxylase protein abundance do not. Additionally, in contrast to certain other enzymes (e.g. phosphoenolpyruvate carboxykinase) it is not possible to mimic diabetes-related alterations in the expression of phenylalanine hydroxylase, in liver cells from normal animals, by simple hormonal manipulation of incubation media. This implies that other additional factors must also contribute to diabetes-related alterations in hepatic enzyme expression.
McDowall
I L
IL
Department of Biochemistry, University of Liverpool, U.K.
Palwala
Y
Y
Fisher
M J
MJ
D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005934:Glucagon / Q000494:pharmacology; D015151:Immunoblotting; D007328:Insulin / Q000494:pharmacology; D007668:Kidney / Q000201:enzymology / Q000378:metabolism; D008099:Liver / Q000201:enzymology* / Q000378:metabolism; D008297:Male; D010651:Phenylalanine Hydroxylase / Q000235:genetics / Q000378:metabolism*; D010730:Phosphoenolpyruvate Carboxylase / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D007328:Insulin; D012333:RNA, Messenger; D003907:Dexamethasone; D005934:Glucagon; D010651:Phenylalanine Hydroxylase; D010730:Phosphoenolpyruvate Carboxylase