title: Effect of Mg2+ on the control of Ca2+ release in skeletal muscle fibres of the toad.
abstract: 1. The effect of myoplasmic Mg2+ on Ca2+ release was examined in mechanically skinned skeletal muscle fibres, in which the normal voltage-sensor control of Ca2+ release is preserved. The voltage sensors could be activated by depolarizing the transverse tubular (T-) system by lowering the [K+] in the bathing solution. 2. Fibres spontaneously contracted when the free [Mg2+] was decreased from 1 to 0.05 mM, with no depolarization or change of total ATP, [Ca2+] or pH (pCa 6.7, 50 microM-EGTA). After such a 'low-Mg2+ response' the sarcoplasmic reticulum (SR) was depleted of Ca2+ and neither depolarization nor caffeine (2 mM) could induce a response, unless the [Mg2+] was raised and the SR reloaded with Ca2+. Exposure to 0.05 mM-Mg2+ at low [Ca2+] (2 mM-free EGTA, pCa greater than 8.7) also induced Ca2+ release and depleted the SR. 3. The response to low [Mg2+] was unaffected by inactivation of the voltage sensors, but was completely blocked by 2 microM-Ruthenium Red indicating that it involved Ca2+ efflux through the normal Ca2+ release channels. 4. In the absence of ATP (and creatine phosphate), complete removal of Mg2+ (i.e. no added Mg2+ with 1 mM-EDTA) did not induce Ca2+ release. Depolarization in the absence of Mg2+ and ATP also did not induce Ca2+ release. 5. Depolarization in 10 mM-Mg2+ (pCa 6.7, 50 microM-EGTA, 8 mM-total ATP) did not produce any response. In the presence of 1 mM-EGTA to chelate most of the released Ca2+, depolarizations in 10 mM-Mg2+ did not noticeably deplete the SR of Ca2+, whereas a single depolarization in 1 mM-Mg2+ (and 1 mM-EGTA) resulted in marked depletion. Depolarization in the presence of D600 and 10 mM-Mg2+ produced use-dependent 'paralysis', indicating that depolarization in 10 mM-Mg2+ did indeed activate the voltage sensors. 6. Depolarization in the presence of 10 mM-Mg2+ and 25 microM-ryanodine neither interfered with the normal voltage control of Ca2+ release nor caused depletion of the Ca2+ in the SR even after returning to 1 mM-Mg2+ for 1 min, indicating that few if any of the release channels had been opened by the depolarization.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Zoology, La Trobe University, Bundoora, Victoria, Australia.
authors.forename: G D
authors.initials: GD
authors.lastname: Lamb
authors.forename: D G
authors.initials: DG
authors.lastname: Stephenson
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002024:Bufo marinus; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism*; D046508:Culture Techniques; D008274:Magnesium / Q000378:metabolism*; D009119:Muscle Contraction / Q000502:physiology; D009132:Muscles / Q000187:drug effects / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D012430:Ruthenium Red / Q000494:pharmacology; D012433:Ryanodine / Q000494:pharmacology; D012519:Sarcoplasmic Reticulum / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D012430:Ruthenium Red; D012433:Ryanodine; D002110:Caffeine; D000255:Adenosine Triphosphate; D008274:Magnesium; D011188:Potassium; D002118:Calcium