pH dependence of hormonal regulation of gluconeogenesis and urea synthesis from glutamine in suspensions of hepatocytes.

Hepatocytes from rats deprived of food for 48 h synthesized glucose and urea from glutamine at a rate which, at pH 7.3, was markedly stimulated (175-250%) by dibutyryl cAMP, phenylephrine, and norepinephrine, in agreement with previous investigators. These effectors also stimulated respiration, elevating ATP production by the amount required for the increase in glucose and urea synthesis. Both the basal and stimulated rates were strongly pH dependent with maxima in the region of pH 7.2-7.6 (urea synthesis) and 7.2-7.5 (glucose synthesis) and declined rapidly on either side of these pH values. The inhibitions at acid and alkaline pH were neither due to lack of energy nor to limitation in glutamine uptake. The intracellular concentrations of aspartate, glutamate, and glutamine were lower at pH 6.7 than at pH 7.3 and were differently affected by dibutyryl cAMP and phenylephrine at the two pH values investigated. When calcium was omitted from the suspending medium, the basal rates of glucose and urea production were decreased as was stimulation by the effectors, phenylephrine completely, and the others partially. The stimulations by phenylephrine and dibutyryl cAMP were additive under all conditions tested. The pattern of metabolite changes indicates that although both effectors stimulated glutaminase and increased supply of aspartate to the argininosuccinate synthetase, dibutyryl cAMP gave greater activation of glutaminase whereas the adrenergic agonists gave greater stimulation of later steps on the biosynthetic pathways. It may be physiologically important than at acid pH both ureagenesis and gluconeogenesis are severely suppressed and cannot be effectively stimulated by the major hormonal regulators of these pathways.