The efflux of magnesium from single crustacean muscle fibres

Abstract

1. In the large single muscle fibres from the barnacle Balanus nubilus, the total fibre Mg concentration was estimated as 15.1 m-mole/kg wet wt., of which about 3-3.5 m-mole/kg wet wt. was extracellular. The diffusible Mg, measured by internal sampling, was 11.5 m-mole/kg wet wt., of which at least half may be complexed to larger diffusible molecules. The free ionized Mg level was estimated as < 5 m-mole/kg wet wt.2. The loss of [(28)Mg]MgCl(2) from both Maia and Balanus muscle fibres following axial micro-injection approximated to first-order kinetics. The maximum rate constant for the loss was 1.51 +/- 0.20 (S.E.) x 10(-5) sec(-1) for Balanus (sixty-seven fibres) and 1.06 +/- 0.46 (S.E.) x 10(-5) sec(-1) for Maia (seven fibres) at 20-25 degrees C.3. The calculated Mg efflux was in the range 6-12 p-mole/cm(2).sec based on this rate constant, assuming isotopic equilibration internally and that the surface area of the fibres approximated to that of a simple cylinder. If account was taken of the area of the cleft system the efflux was reduced by about fifteen times.4. The diffusion coefficient for injected (28)Mg was estimated as 2-3 x 10(-6) cm(2) sec(-1), about half the value in free solution. Injections of 2 M-MgCl(2) or 200 mM-EDTA subsequent to the injection of the isotope caused about a 30% reduction in the tracer efflux.5. External application of salines containing 100 mM-Ca or Mg caused a rapid but reversible inhibition of the magnesium efflux. Similar effects were observed with salines containing 32 mM-Co or Mn chlorides or 1-2 mM-La or Gd chlorides. Polyarginine (200 mug/ml.) had no effect.6. The Mg efflux had a Q(10) of 3-4 over the temperature range of about 5-20 degrees C. It was irreversibly inhibited by the sulphydryl reagent NEM (1 mM), but PCMBS (0.2-2 mM) had no effect. Contractile agents (5 mM caffeine or 200 mM-K salines) and a variety of inhibitors of ion movement or active transport had no appreciable effect on the Mg efflux. Lowering the pH of the saline from 7 to 5 produced a 70% reduction in the efflux which was reversible over short periods of application.7. Replacement of external Na, but not Ca or Mg, with Li, choline or sucrose caused a rapid and partially reversible reduction of the Mg efflux, but increasing the internal Na by micro-injection in zero Na salines had no consistent effect. It is suggested that the extrusion of Mg from these muscle cells is largely dependent upon the inward movement of Na.