Metabolism in the cytosol of intact isolated cattle rod outer segments as indicator for cytosolic calcium and magnesium ions.

The metabolism of the chromophore of rhodopsin in the cytosol compartment of isolated intact cattle rod outer segments was used as an indicator for changes of the cytosolic Mg2+ and Ca2+ concentration upon changes of the external Mg2+ and Ca2+ concentration. The reduction of retinal to retinol upon photolysis of rhodopsin in situ in intact rod outer segments was critically dependent on the availability of cytosolic Mg2+. The latter is necessary as chelator of endogenous adenosine 5'-triphosphate (ATP). Lowering the cytosolic Ca2+ concentration beneath 10(-7) M resulted in an inhibition of the rate of retinol formation. This is presumably due to a light-activated process, which competes with retinol formation for the supply of high-energy phosphate from a common pool. These results led to the following conclusions. Changes of the external Mg2+ concentration are only followed by substantial changes of the cytosolic Mg2+ concentration when the ionophore A23187 is present. Changes of the external Ca2+ concentration are followed by parallel changes of the cytosolic Ca2+ concentration either when external Na+ is present or in the presence of A23187. Li+ and K+ could not substitute for Na+ in the former case, but K+ diminished the effectivity of Na+ at low Na+ concentrations and enhanced it at high Na+ concentrations. It is concluded that the control of cytosolic Ca2+ concentration in isolated intact rod outer segments is predominantly provided for by Na-Ca exchange, i.e., by coupled fluxes.