A convenient HPLC assay, which allows for the simultaneous measurement in extracellular fluids of 6-mercaptopurine and four of its metabolites, 6-thioguanine, 6-mercaptopurine riboside, 6-thioxanthine and 6-thiouric acid is described. Solid phase extraction allows for the clean isolation of analytes from plasma, urine or cerebrospinal fluid. The simultaneous determination of 6-mercaptopurine and some of its major metabolites in extracellular fluids may contribute to the monitoring of patient compliance, bioavailability, and individual variation in metabolism and absorption.

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We investigated physiological interactions between alpha 2-adrenoceptors and parathyroid hormone (PTH) in the isolated buffer-perfused kidney. PTH infusion (10nM) caused a rapid and significant rise in cAMP excretion which diminished despite continued hormone infusion. PTH also caused a delayed phosphaturia which peaked 20-30 minutes following the start of PTH infusion. alpha 2-Adrenoceptor stimulation with epinephrine (28nM) diminished PTH-stimulated cAMP accumulation by 68-71% (p less than 0.05) but had no effect on PTH-induced phosphaturia. None of the experimental interventions affected renal hemodynamics. In additional studies, we used lithium (1mM) as a marker of proximal tubular sodium transport. PTH caused a rapid rise in lithium excretion which was temporally distinct (maximum response in 0-10 minutes) from the phosphaturia. alpha 2-Adrenoceptor stimulation completely blocked the inhibitory effect of PTH on lithium reabsorption. These results suggest that alpha 2-adrenoceptors regulate the stimulatory effect of PTH at proximal tubular adenylate cyclase. However, alpha 2-adrenoceptors play no role in phosphate transport in this segment. alpha 2-Adrenoceptor stimulation reverses PTH-induced lithium excretion, suggesting physiological antagonism in the proximal tubule, most likely involving Na/H exchange.

The putative role of the inhibitory guanine nucleotide binding protein (Gi) in modulating the renal response to vasopressin was investigated using islet activating protein (IAP). IAP treatment in rats in vivo abolished the capacity of alpha 2-adrenoceptors to reverse vasopressin-induced adenosine 3',5'-cyclic monophosphate (cAMP) accumulation in microdissected cortical collecting tubule (CCT) segments. IAP pretreatment also caused a marked upward shift in the dose-response curve of vasopressin (10(-10) to 10(-4) M)-induced cAMP accumulation. Augmentation of the response to vasopressin in rat CCT was dependent on the in vivo dose of IAP and paralleled the loss in alpha 2-adrenoceptor responsiveness. In the isolated perfused kidney the antinatriuretic and antidiuretic effects of the V2-receptor agonist desamino-8-D-arginine vasopressin (DDAVP) (1 pM) were enhanced following IAP pretreatment. alpha 2-Adrenoceptor stimulation (30 nM epinephrine) inhibited the renal effects of DDAVP (1 pM) in kidneys from control but not IAP-pretreated rats. Interestingly, IAP pretreatment alone caused increased urine flow rate and enhanced excretion of sodium and chloride without affecting potassium excretion or renal hemodynamics in vitro. Our results suggest that an IAP substrate, probably Gi, 1) is required for signal transduction by renal alpha 2-adrenoceptors, 2) may tonically modulate the response to vasopressin in the CCT but not of parathyroid hormone in the proximal convoluted tubule, and 3) participates in renal water and electrolyte reabsorption independent of exogenous adenylate cyclase stimulation.