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Photokinetic assay of pyruvate in the islets of langerhans using bacterial luciferase

✍ Scribed by Ambjörn Ågren; Christian Berne; Sven E. Brolin

Publisher
Elsevier Science
Year
1977
Tongue
English
Weight
359 KB
Volume
78
Category
Article
ISSN
0003-2697

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✦ Synopsis

A method is described determining the pyruvate content in l-10 pg of lyophilized tissue. This represents a range of weight much below that of fine-needle biopsies. NAD+, generated through the enzymatic conversion of pyruvate to lactate, is reduced to NADH which is then assayed by its ability to induce light emission with h&erase extracts ofAchromobacter$scherii.

The sensitivity of the method is sufficient to determine 0.5 pmol of pyruvate. Values obtained by this method in lyophilized liver agree well with those obtained by spectrophotometric techniques. In applications of the method, it was found that the islet concentration of pyruvate was greater than in the exocrine pancreas but smaller than in the liver. The pyruvate content in tissues of the obese hyperglycemic mice was greater than in their lean litter mates, which may be explained by a more rapid rate of glucose utilization due to their hyperglycemia.

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