Simultaneous multicompartment intracellular Ca2+ measurements in the perfused heart using 19F NMR spectroscopy.
Although Ca2+ transport regulation at subcellular organelles is of great interest, only limited methodology has been available for measuring organellar [Ca2+] levels. The present study employs the 19F NMR resonance frequency of 4F-BAPTA to measure free [Ca2+]. In 4F-BAPTA loaded perfused rabbit hearts, two 19F NMR resonances were clearly observed. The frequency of one was consistent with cytosolic [Ca2+] levels. Responses to agents that after sarcoplasmic reticulum function identified the other resonance as originating from that organelle. The experiments demonstrate the utility of NMR shift indicator methodology in obtaining simultaneous multi-compartment intracellular [Ca2+] measurements and in enabling organellar [Ca2+] measurements to be made from within intact living tissue.
Yanagida
S
S
Department of Medicine, University of Alabama at Birmingham 35294-0007, USA.
Luo
C S
CS
Balschi
J A
JA
Pohost
G M
GM
Pike
M M
MM
D000818:Animals; D001693:Biological Transport, Active; D002118:Calcium / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004533:Egtazic Acid / Q000031:analogs & derivatives; D005461:Fluorine; D006321:Heart; D007202:Indicators and Reagents; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009206:Myocardium / Q000378:metabolism*; D010477:Perfusion; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000378:metabolism
D007202:Indicators and Reagents; D005461:Fluorine; D004533:Egtazic Acid; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium