Calcium gradients during excitation-contraction coupling in cat atrial myocytes.
1. Confocal microscopy in combination with the calcium-sensitive fluorescent probe fluo-3 was used to study spatial aspects of intracellular Ca2+ signals during excitation-contraction coupling in isolated atrial myocytes from cat heart. 2. Imaging of [Ca2+]i transients evoked by electrical stimulation revealed that Ca2+ release started at the periphery and subsequently spread towards the centre of the myocyte. 3. Blocking sarcoplasmic reticulum (SR) Ca2+ release with 50 microM ryanodine unmasked spatial inhomogeneities in the [Ca2+]i was higher in the periphery than in central regions of the myocyte. 4. Positive (or negative) staircase or postrest potentiation of the 'whole-cell' [Ca2+] transients were paralleled by characteristic changes in the spatial profile of the [Ca2+]i signal. With low SR Ca2+ load [Ca2+]i transients in the subsarcolemmal space were small and no Ca2+ release in the centre of the cell was observed. Loading of the SR increased subsarcolemmal [Ca2+]i transient amplitude and subsequently triggered further release in more central regions of the cell. 5. Spontaneous Ca2+ release from functional SR units, i.e. Ca2+ sparks, occurred at higher frequency in the subsarcolemmal space than in more central regions of the myocyte. 6. Visualization of the surface membrane using the membrane-selective dye Di-8-ANEPPS demonstrated that transverse tubules (t-tubules) were absent in atrial cells. 7. It is concluded that in atrial myocytes voltage-dependent Ca2+ entry triggers Ca2+ release from peripheral coupling SR that subsequently induces further Ca2+ release from stores in more central regions of the myocyte. Spreading of Ca2+ release from the cell periphery to the centre accounts for [Ca2+]i gradients underlying the whole-cell [Ca2+]i transient. The finding that cat atrial myocytes lack t-tubules demonstrates the functional importance of Ca2+ release from extended junctional (corbular) SR in these cells.
Hüser
J
J
Department of Physiology, Stritch School of Medicine, Loyola University Chicago, Maywood, IL 60153, USA.
Lipsius
S L
SL
Blatter
L A
LA
D000818:Animals; D016275:Atrial Function; D002118:Calcium / Q000502:physiology*; D002415:Cats; D005260:Female; D006325:Heart Atria / Q000648:ultrastructure*; D008297:Male; D008564:Membrane Potentials / Q000502:physiology*; D018613:Microscopy, Confocal; D009119:Muscle Contraction / Q000502:physiology*
D002118:Calcium