Substrate oxidation was assessed by measuring 14CO2 production from 14C-labeled substrates in proximal convoluted tubules (PCT), medullary (MTAL), and cortical (CTAL) thick ascending limb of Henle, nephron segments rich in mitochondria and characterized by active solute transport. PCT, MTAL, and CTAL were dissected from the outer cortex, outer medulla, and the medullary rays of the cortex, respectively, of collagenase-treated rat kidney slices. Tubules were incubated at 37 degrees C in 150 microliters of Krebs-Ringer-bicarbonate buffer (pH, 7.4) with 14C-labeled substrate. 14CO2 production was linear up to 4 and 2 hours in PCT and MTAL, respectively. Freeze-thawing of the tubules markedly decreased 14CO2 production, and the addition of cyanide completely abolished it. The PCT demonstrated marked 14CO2 production from labeled succinate, 2-oxoglutarate, glutamate, glutamine, and malate (approximately 10 to 45 pmoles/mm/hr) and moderate 14CO/ production from citrate (approximately 3 pmoles/ml/hr). Little 14CO2 was released from labeled glucose and lactate in PCT. These results are consistent with the existence of gluconeogenesis in this nephron segment. By contrast, MTAL and CTAL oxidized glucose, 2-oxoglutarate, lactate, glutamate, and glutamine, but not malate, succinate, and citrate. The pentose shunt pathway accounted for approximately half of the 14CO2 produced from 1-14C glucose in MTAL and CTAL. Palmitate oxidation occurred in MTAL and CTAL but minimally in PCT. The results demonstrate a distinct pattern of substrate oxidation in PCT, MTAL, and CTAL where oxidative metabolism is critical to support active solute transport.

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Parathyroid hormone (PTH: synthetic bovine, amino terminus 1-34 amino acids) demonstrates a positive inotropic action on the isolated papillary muscle of the rat heart. The effect was evident at PTH concentration of 10(-12)M, and the maximum inotropic effect occurred with PTH concentrations greater than 10(-11)M. Biologically inactive PTH (PTH treated with H2O2) was without effect. The inotropic effect of PTH was partially blocked by propranolol and also suppressed in the papillary muscle of the rat pretreated with reserpine. Methoxyverapamil completely blocked the inotropic action of PTH. PTH was without effects on adenylate cyclase activity of the myocardium. Results show the presence of an inotropic action of PTH in vitro and suggest that this action of PTH is partially mediated by releasing the endogenous myocardial norepinephrine which exerts a positive inotropic effect via beta-adrenergic stimulation and by an increase in Ca++ influx across plasma membranes, but independent of adenylate cyclase activation. The inotropic action of PTH may be of significance in normal cardiac function.

A B S T R A C T To investigate a possible action of insulin on the glomerulus, the binding 1251-insulin to the isolated glomeruli prepared from rat kidney was examined. When incubated at 22°C, '251-insulin binding proceeded with time and reached a steady state at 45 min at which time nonspecific binding was <25% of total binding. A small fraction of '25I-insulin was degraded during incubation. This binding was specific to insulin in that it was inhibited by unlabeled porcine and beef insulins and to a lesser extent by porcine proinsulin and desalanine-desasparagine insulin, but not by glucagon, parathyroid hormone, vasopressin, calcitonin, and angiotensin II. Increasing concentrations of nonlabeled insulin displaced 1251-insulin binding in a dose-dependent fashion. Scatchard plot of the data was curvilinear consistent with either two classes of receptors with different affinities or a single class of receptors that demonstrate negative cooperativity. The addition of excess nonlabeled insulin to the glomeruli preincubated with 1251-insulin resulted in a rapid dissociation of -70% of bound I251-insulin. Insulin decreased the increments in glomerular cyclic AMP levels by epinephrine and by prostaglandin E2, but not those by histamine. These data showed the presence of specific insulin receptors in the glomeruli, and that insulin action may be, at least in part, through modulation of glomerular cyclic AMP concentrations. Such action of insulin may underlie the alteration in glo-

Effects of parathyroid hormone (PTH), calcitonin (CT), vasopressin (VP), and glucagon (GL) on adenylate cyclase activity and cyclic AMP (cAMP) levels in isolated cortical thick ascending limbs of Henle's loop (CTAL) of the rat kidney were examined. PTH, CT, and VP each stimulated adenylate cyclase of this nephron segment in a dose-dependent manner. Stimulation of the enzyme activity was greatest with a maximal dose of PTH and least with VP. With maximal doses, the effects of PTH and CT were not additive; whether or not the effects of maximal doses of VP and PTH or CT were additive was not clear. All three hormones increase cAMP in intact CTAL ina dose-dependent manner. Maximal doses of PTH, CT, VP, and GL resulted in comparable rises in cell cAMP, and there was no additive effect. These data suggest that PTH and CT may stimulate the same adenylate cyclase moieties, whereas VP may stimulate distinct enzyme moieties, and that these three peptide hormones as well as GL definitely act on the same cell group in rat CTAL. Thus, it is possible that these hormones may induce qualitatively similar effects on CTAL functions if such effects are mediated by cAMP.