TY - JOUR
T1 - Mitochondrial Ca2+ transients in cardiac myocytes during the excitation-contraction cycle
T2 - Effects of pacing and hormonal stimulation
AU - Ohata, Hisayuki
AU - Chacon, Enrique
AU - Tesfai, Samuel A.
AU - Harper, Ian S.
AU - Herman, Brian
AU - Lemasters, John J.
PY - 1998/1/1
Y1 - 1998/1/1
N2 - Using laser scanning confocal microscopy, our objective was to measure mitochondrial, nuclear, and cytosolic free ionized Ca2+ in adult rabbit cardiac myocytes loaded with Ca2+ -indicating fluorophores. When myocytes were loaded with Flou 3 at 37°C, the fluorophore was loaded extensively into the cytosol and nucleus, but poorly into mitochondria, and Fluo 3 fluorescence transients after field stimulation were confined to the cytosol and nucleus. In contrast, after loading at 4°C, Fluo 3 also entered mitochondria, and large transients of mitochondrial Fluo 3 fluorescence then occurred after stimulation. Isoproterenol (1 μM) increased the magnitude of Ca2+ transients and their subsequent rate decay, an effect more marked in the cytosol and nucleus than in the mitochondria. As pacing frequency was increased from 0.5 to 2 Hz, diastolic mitochondrial Ca2+ rose markedly in the absence but not in the presence of isoproterenol. Resting Ca2+ estimated by Indo 1 ratio imaging using UV/visible laser scanning confocal microscopy was about 200 nM in all compartments. During field stimulation, Ca2+ transiently increased to 671, 522, and 487 nM in cytosol, interfibrillar mitochondria, and perinuclear mitochondria, respectively. Isoproterenol increased these respective peak values to 1280, 750, and 573 nM. These results were consistent with those obtained in Fluo 3 experiments. We conclude that rapid mitochondrial Ca2+ transients occur during excitation-contraction coupling in adult rabbit cardiac myocytes, which may be important in matching mitochondrial metabolism to myocardial ATP demand during changes in cardiac output.
AB - Using laser scanning confocal microscopy, our objective was to measure mitochondrial, nuclear, and cytosolic free ionized Ca2+ in adult rabbit cardiac myocytes loaded with Ca2+ -indicating fluorophores. When myocytes were loaded with Flou 3 at 37°C, the fluorophore was loaded extensively into the cytosol and nucleus, but poorly into mitochondria, and Fluo 3 fluorescence transients after field stimulation were confined to the cytosol and nucleus. In contrast, after loading at 4°C, Fluo 3 also entered mitochondria, and large transients of mitochondrial Fluo 3 fluorescence then occurred after stimulation. Isoproterenol (1 μM) increased the magnitude of Ca2+ transients and their subsequent rate decay, an effect more marked in the cytosol and nucleus than in the mitochondria. As pacing frequency was increased from 0.5 to 2 Hz, diastolic mitochondrial Ca2+ rose markedly in the absence but not in the presence of isoproterenol. Resting Ca2+ estimated by Indo 1 ratio imaging using UV/visible laser scanning confocal microscopy was about 200 nM in all compartments. During field stimulation, Ca2+ transiently increased to 671, 522, and 487 nM in cytosol, interfibrillar mitochondria, and perinuclear mitochondria, respectively. Isoproterenol increased these respective peak values to 1280, 750, and 573 nM. These results were consistent with those obtained in Fluo 3 experiments. We conclude that rapid mitochondrial Ca2+ transients occur during excitation-contraction coupling in adult rabbit cardiac myocytes, which may be important in matching mitochondrial metabolism to myocardial ATP demand during changes in cardiac output.
KW - Calcium
KW - Cardiac myocytes
KW - Confocal microscopy
KW - Fluo 3
KW - Indo 1
KW - Isoproterenol
KW - Mitochondria
UR - http://www.scopus.com/inward/record.url?scp=0031593710&partnerID=8YFLogxK
U2 - 10.1023/A:1020588618496
DO - 10.1023/A:1020588618496
M3 - Article
C2 - 9733088
AN - SCOPUS:0031593710
SN - 0145-479X
VL - 30
SP - 207
EP - 222
JO - Journal of Bioenergetics and Biomembranes
JF - Journal of Bioenergetics and Biomembranes
IS - 3
ER -