Abstract
Detubulation of rat ventricular myocytes has been used to investigate
the role of the t-tubules in Ca$^2+$ cycling during excitation-contraction
coupling in rat ventricular myocytes. Ca$^2+$ was monitored using
fluo-3 and confocal microscopy. In control myocytes, electrical stimulation
caused a spatially uniform increase in intracellular Ca$^2+$
across the cell width. After detubulation, Ca$^2+$ rose initially
at the cell periphery and then propagated into the center of the
cell. Application of caffeine to control myocytes resulted in a rapid
and uniform increase of intracellular Ca$^2+$; the distribution
and amplitude of this increase was the same in detubulated myocytes,
although its decline was slower. On application of caffeine to control
cells, there was a large, rapid, and transient rise in extracellular
Ca$^2+$ as Ca$^2+$ was extruded from the cell; this rise
was significantly smaller in detubulated cells, and the remaining
increase was blocked by the sarcolemmal Ca$^2+$ ATPase inhibitor
carboxyeosin. The treatment used to produce detubulation had no significant
effect on Ca$^2+$ efflux in atrial cells, which lack t-tubules.
Detubulation of ventricular myocytes also resulted in loss of Na$^+$-Ca$^2+$
exchange current, although the density of the fast Na$^+$ current
was unaltered. It is concluded that Na$^+$-Ca$^2+$ exchange
function, and hence Ca$^2+$ efflux by this mechanism, is concentrated
in the t-tubules, and that the concentration of Ca$^2+$ flux
pathways in the t-tubules is important in producing a uniform increase
in intracellular Ca$^2+$ on stimulation.
- 12193458
- action
- adenosinetriphosphatase,
- agents,
- animals,
- atria,
- caffeine,
- calcium,
- capacitance,
- cardiotonic
- cell
- compounds,
- confocal,
- dyes,
- electric
- electrophysiology,
- enzyme
- exchanger,
- fibers,
- fluid,
- fluorescent
- formamides,
- gov't,
- heart
- inhibitors,
- intracellular
- isoproterenol,
- membrane
- microscopy,
- microtubules,
- muscle
- myocardium,
- nickel,
- non-u.s.
- patch-clamp
- potentials,
- pyridinium
- rats,
- research
- sarcolemma,
- separation,
- sodium-calcium
- stimulation,
- structures,
- support,
- techniques,
- ventricles,
- wistar,
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