A sensitive radioisotopic assay of myo-inositol: Its application to rat pancreatic islets |
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Institution: | 1. Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel;2. Department of Neurobiology, Weizmann Institute of Science, Rehovot 76100, Israel;3. Department of Veterinary Resources, Weizmann Institute of Science, Rehovot 76100, Israel |
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Abstract: | A radioisotopic procedure for the assay of myo-inositol is presented. It is based on the generation of NADH from NAD+ in the reaction catalyzed by myo-inositol dehydrogenase and the subsequent NADH-dependent conversion of 2-U-14C]ketoglutarate to 14C-labeled l-glutamate in the reaction catalyzed by glutamate dehydrogenase. This method was applied to the measurement of myo-inositol in rat pancreatic islets. The myo-inositol islet content was decreased when the animals were fed a diet deprived of myo-inositol. When incubated in the absence of exogenous d-glucose, pancreatic islets, like parotid cells, released myo-inositol in the incubation medium. Over 90 min of incubation, a rise in extracellular d-glucose concentration increased the myo-inositol islet content, which was decreased, however, after incubation in the presence of carbamylcholine. These findings indicate that the myo-inositol content of islets is affected by nutritional and other environmental factors. |
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