A cytoplasmic component of pyridine nucleotide fluorescence in rat diaphragm: evidence from comparisons with flavoprotein fluorescence

Abstract

Pyridine nucleotide (PN) and flavoprotein (Fp) fluorescence were monitored in the isolated intact rat diaphragm. A substantial increase in PN fluorescence occurred when N₂ replaced O₂ in glucose medium. This response was much reduced in pyruvate medium and/or by pretreatment with iodoacetic acid (IAA). The anaerobic levels of Fp fluorescence were less affected by substrate and IAA. Substitution of glucose by pyruvate did not alter the PN fluorescence of the resting aerobic tissue, but increased Fp fluorescence. After a tetanus with glucose present the PN of the anaerobic muscle, but not the Fp underwent a substantial transient oxidation. This oxidation was absent in pyruvate medium. It is concluded that a cytoplasmic component of the PN fluorescence is present in skeletal muscle. The levels of Fp fluorescence in the resting and contracting aerobic tissue supplied with pyruvate suggest that the resting tissue respiration was ADP limited. On this basis the level of PN fluorescence in the aerobic resting state was less than expected; the source of the PN fluorescence was both mitochondrial and cytoplasmic.