Changes of Cytosolic Ca2+ under Metabolic Inhibition in Isolated Rat Ventricular Myocytes
The Korean Journal of Physiology and Pharmacology
; : 291-298, 2005.
Article
em En
| WPRIM
| ID: wpr-728717
Biblioteca responsável:
WPRO
ABSTRACT
To characterize cytosolic Ca2+ fluctuations under metabolic inhibition, rat ventricular myocytes were exposed to 200microM 2, 4-dinitrophenol (DNP), and mitochondrial Ca2+, mitochondrial membrane potential (delta psi m), and cytosolic Ca2+ were measured, using Rhod-2 AM, TMRE, and Fluo-4 AM fluorescent dyes, respectively, by Laser Scanning Confocal Microscopy (LSCM). Furthermore, the role of sarcolemmal Na+/Ca2+ exchange (NCX) in cytosolic Ca2+ efflux was studied in KB-R7943 and Na+-free normal Tyrode's solution (143 mM LiCl ). When DNP was applied to cells loaded with Fluo-4 AM, Fluo-4 AM fluorescence intensity initially increased by 70+/-10% within 70+/-10 s, and later by 400+/-200% at 850+/-46 s. Fluorescence intensity of both Rhod-2 AM and TMRE were initially decreased by DNP, coincident with the initial increase of Fluo-4 AM fluorescence intensity. When sarcoplasmic reticulum (SR) Ca2+ was depleted by 1microM thapsigargin plus 10microM ryanodine, the initial increase of Fluo-4 AM fluorescence intensity was unaffected, however, the subsequent progressive increase was abolished. KB-R7943 delayed both the first and the second phases of cytosolic Ca2+ overload, while Na+-free solution accelerated the second. The above results suggest that: 1) the initial rise in cytosolic Ca2+ under DNP results from mitochondrial depolarization; 2) the secondary increase is caused by progressive Ca2+ release from SR; 3) NCX plays an important role in transient cytosolic Ca2+ shifts under metabolic inhibition with DNP.
Palavras-chave
Texto completo:
1
Base de dados:
WPRIM
Assunto principal:
Rianodina
/
Retículo Sarcoplasmático
/
Cafeína
/
Microscopia Confocal
/
Tapsigargina
/
Citosol
/
Células Musculares
/
Potencial da Membrana Mitocondrial
/
Fluorescência
/
Corantes Fluorescentes
Limite:
Animals
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article