Osmotic changes and transsarcolemmal ion transport during total ischaemia of isolated rat ventricular myocytes

Transsarcolemmal water and ion movement during 1, 7.5, 15, and 30 min of total ischaemia was studied in suspensions of isolated rat ventricular myocytes, with a control ratio of about 1 of intracellular volume (ICV) to extracellular volume (ECV). In this preparation, contrary to the intact heart: 1) There is no external exchange of matter, 2) the sum of ICV and ECV remains constant and 3) ECV is homogeneous; no separate interstitial and intravascular compartments are present and no extracellular metabolite or ion gradients develop as may occur in the intact heart. We demonstrate that: 1) It is possible to make an ischaemic preparation of isolated myocytes with a procedure which causes only minimal mechanical damage to intact myocytes. The preparation allows measurement of ECV with the non-cardiac enzyme alpha-amylase as a macromolecular extracellular marker. 2) The time course of change of metabolites relevant to energy metabolism (creatinephosphate (CrP), creatine (Cr), ATP, ADP, inorganic phosphate P(i) and lactate) is similar to that in the intact heart. 3) ECV has decreased and ICV increased by about 20% after 30 min of ischaemia. 4) Extracellular [Na+], [K+], [Cl-], and [P(i)] increase, but not in proportion to the decrease of ECV. There is net efflux of K+, P(i), H+, and lactate-; efflux of K+ and P(i) is quantitatively much less than influx of Na+ and Cl-. 5) Measured extracellular osmolality has increased with up to 70 mOsm/l after 30 min of ischaemia. The increase of extracellular [lactate-], [Na+], [K+], [Cl-], [P(i)] and the decrease of [glucose] account for the change of osmolality measured. 6) Summation of the electrical charges associated with measured increase of extracellular [lactate-], [Na+], [K+], [Cl-], [P(i)] shows a surplus of negative charge, which almost equals extracellular [lactate-], suggesting an equally large increase of osmotically inactive H+ as the compensatory ion. 7) Blockade of anaerobic metabolism with iodoacetic acid (IAA) reduces efflux of lactate and P(i) but greatly amplifies influx of sodium and chloride and efflux of potassium.