Tonic contraction of canine gastric muscle during long-lasting calcium removal and its dependence on magnesium.

Abstract

1. Tonic contractions induced by acetylcholine (ACh) in canine gastric fundus preparations were shown to persist during long-lasting exposure to Ca2+-free solution containing EGTA (1 mmol/l). These EGTA-resistant contractions amounted to up to more than 50% of maximal ACh-control responses in physiological salt solution. They could be evoked repeatedly for more than 20 h without reduction in size, each contraction lasting as long as ACh was present. 2. During prolonged exposure to Ca2+-free solution at normal Mg2+ concentration ([Mg2+]O = 1.2 mmol/l), the preparations exhibited a slowly developing contracture (elevation of the baseline of contraction), which was particularly pronounced in strips taken from the circular layer of the muscular wall (44% of control ACh-maximum after 4 h in Ca2+-free solution). Contracture could be suppressed either by increasing [Mg2+]O to 6-10 mmol/l or by depolarizing the cell membrane (replacement of external Na+ by K+). However, contracture also developed if, at physiological [Na+]O and [K+]O, [Mg2+]O was further increased to 50 mmol/l. 3. The combined effects of [Mg2+]O and membrane potential suggest that contracture is caused by a gain of Mg2+ by the cells. This conclusion is based on the assumption that (a) the cytoplasmic Mg2+ concentration is determined by the transmembrane electrochemical gradient acting on Mg2+, the magnesium permeability of the cell membrane (PMg) and an active extrusion mechanism, and that (b) Ca2+ removal leads to an increase of PMg which is (partly) prevented by an appropriate increase of [Mg2+]O. 4. 45Ca efflux experiments, performed at [Mg2+]O = 10 mmol/l to avoid interference of ACh responses with contracture, showed that the cellular 45Ca content decreased from some 200 mumol/kg wet wt. to less than 10 mumol/kg wet wt. within 10-20 h in Ca2+-free solution. Activations by ACh did not produce any detectable increase in 45Ca efflux. 5. The calcium ionophore A23187 (10(-5) mol/l), applied in order to increase the calcium permeability of the cell membrane, did not reduce the EGTA-resistant contractions. 6. Experimental procedures conducted to replace calcium within intracellular stores by strontium or barium did not affect the EGTA-resistant ACh response. However, prior uptake of manganese by the cells had an amplifying effect. 7. Caffeine (30 mmol/l) failed to produce contraction in Ca2+-free solution, whereas ACh evoked contractile responses, both in the presence of and after application of caffeine.(ABSTRACT TRUNCATED AT 400 WORDS)