A new method to measure cardiac inositol levels in intact animals

Abstract

Inositol levels have been studied in cellular cultures and recently by perfusion of isolated hearts. The study was aimed to assess inositol turnover in rabbit hearts from intact animals. Thirty rabbits were injected i.v. three times (every 12 hr) with 25 μCi/kg of myo-3H-inositol. The rabbits 12 hr after the last injection were killed and the hearts perfused according to Langerdorff technique. Systolic and diastolic ventricular pressures (SVP, DVP), dp dt , and coronary flow (CFl) were measured. The hearts ( n = 14) were perfused under aerobic conditions and 16 hearts under ischemic conditions for 30 min. In addition, 5 hearts were perfused under aerobic conditions for 10 min, and 6 hearts were perfused under ischemic conditions for 10 min. Samples of myocardial tissue were taken from both groups at the end of 10-min and 30-min period of perfusion, and cAMP and inositol phosphates were assayed. The hearts subjected to ischaemia showed changes of cAMP and 3H-inositol. The cAMP was higher in the ischaemic (10 min and 30 min) than the control hearts, 0.22 ± 0.09 and 0.21 ± 0.08 versus 0.41 ± 0.12 and 0.49 ± 0.11 pmol 10 6 cells, respectively ( p < .05, p < .001. The inositol trisphosphate was higher in control than ischemic hearts (10 min, 30 min), 0.42 ± 0.02 and 0.39 ± 0.01 versus 0.31 ± 0.01 and 0.23 ± 0.02 (percent of radioactivity) respectively, p < .001. Our data suggest that 3H-inositol may be studied by i.v. administration to intact animals. The ischemia was performed to verify the validity of this new technique.