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The effect of electrical stimulation and ouabain on the uptake and efflux of L-[U-14C] valine in chopped tissue from guinea-pig cerebral cortex.

ABSTRACT: 1. Chopped tissue from guinea-pig cerebral cortex carried out an energy-dependent accumulation of l-[(14)C]valine. 2. The uptake was dependent on the extracellular concentration of Na(+) and was markedly inhibited by ouabain (20mum). The extent of the inhibition of uptake by ouabain was also Na(+)-dependent. 3. The accumulation of labelled valine was not directly dependent on the ATP and creatine phosphate contents of the slices. 4. Electrical stimulation increased the rate of [(14)C]valine uptake at first but ultimately led to a net loss of the label so that the amount of label present in the tissue was lower than in the controls. 5. The rate of loss of label during prolonged stimulation was dependent on the extracellular concentration of Na(+). 6. The efflux of labelled valine from slices preloaded at 164mm-Na(+) was studied at 164, 80 and 40mm-Na(+) with and without electrical stimulation or ouabain. 7. Lowering the Na(+) concentration or adding ouabain increased the rate of efflux. 8. Electrical stimulation had little effect on the rate of efflux at first but ultimately led to a more complete loss of label from the tissue than occurred in the control. A kinetic analysis of the efflux curves was attempted.

SUBMITTER: Jones CT

PROVIDER: S-EPMC1179290 | biostudies-other | 1970 Aug

REPOSITORIES: biostudies-other

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The effect of electrical stimulation on the incorporation of L-[U-14C] valine into the protein of chopped tissue from guinea-pig cerebral cortex.

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| S-EPMC1179289 | biostudies-other

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| S-EPMC1178048 | biostudies-other

Mechanisms of electrical remodeling in lipotoxic guinea pig heart.

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Project description:Inhibition of receptor-coupled adenylate cyclase by hormones is proposed to be associated with GTP hydrolysis. Since adenosine inhibits cerebral-cortical adenylate cyclase via A1 adenosine receptors, the present study attempts to verify this mechanism for A1-selective adenosine derivatives. In guinea-pig cortical membranes N6-(phenylisopropyl)adenosine (PIA) increased the Vmax. of the low-Km GTPase, with an EC50 (concentration causing 50% of maximal stimulation) of about 0.1 microM, and the stimulatory effect was competitively antagonized by 5 microM-8-phenyltheophylline. The rank order of potency of the stereoisomers of PIA and of 5-(N-ethylcarboxamido)adenosine (NECA) to stimulate GTPase correlated with their ability to inhibit adenylate cyclase activity (R-PIA greater than NECA greater than S-PIA). Competition binding studies with (-)-N6- ([125I]iodo-4-hydroxyphenylisopropyl)adenosine suggest that adenylyl imidodiphosphate (p[NH]ppA), an essential component of the GTPase assay system, is a more potent A1-receptor agonist than ATP, with an IC50 (concentration giving half-maximal displacement of radioligand binding) of 7.9 microM. On the basis of the p[NH]ppA concentration used in the GTPase assay (1.25 mM), enzyme stimulation by adenosine seems to be highly underestimated. Nevertheless, adenosine-induced GTP hydrolysis reflects an increased turnover of guanine nucleotides at the Ni regulatory site and appears to be a crucial step in the sequence of events processing the inhibitory signal to adenylate cyclase.

| S-EPMC1152908 | biostudies-other

The subcellular distribution and properties of hexokinases in the guinea-pig cerebral cortex.

Project description:1. Hexokinase activities were estimated in primary subcellular fractions from guinea-pig cerebral cortex and in sucrose-density-gradient subfractions of the mitochondrial and microsomal fractions. 2. Appreciable activities were observed in mitochondrial, microsomal and soluble fractions. The activity in the mitochondrial fraction was associated with the mitochondria rather than with myelin or nerve endings and that in the microsomal fraction was associated with membrane fragments. 3. Most of the mitochondrial activity was extracted in soluble form by osmotic ;shock'. The activity of the mitochondrial extract differed from the soluble activity in kinetic properties and in electrophoretic behaviour. 4. No evidence was obtained for the presence of a high-K(m) glucokinase in the brain. 5. The results are discussed in terms of relevance to considerations of glucose utilization by the brain.

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