PulseExploreJournal ClubDebatesTrendingResearchersJournals
Instagram
HomeExploreJournal ClubTrending
Synapse
⌘+K
Synapse
October 1, 1993Cardiovascular Research167 citations

The relative contributions of different intracellular and sarcolemmal systems to relaxation in rat ventricular myocytes

View Full Paper
NNNicholas M. NegrettiSOStephen O’NeillDEDavid Eisner

Key Result

Calcium removal in isolated rat ventricular myocytes was primarily mediated by the sarcoplasmic reticulum (87%), followed by Na-Ca exchange (8.7%), sarcolemmal Ca-ATPase (2.6%), and mitochondria (1.7%).

Key Points

  • To quantify the relative contributions of intracellular organelles and sarcolemmal transport systems to the relaxation of systolic intracellular calcium transients in rat cardiac cells.
  • Isolated rat ventricular myocytes were loaded with the fluorescent indicator indo-1 to measure intracellular calcium concentration ([Ca2+]i).
  • Specific pathways were sequentially evaluated using caffeine to deplete the sarcoplasmic reticulum, nickel (Ni2+) to inhibit Na-Ca exchange, elevated extracellular calcium to inhibit sarcolemmal Ca-ATPase, and FCCP plus oligomycin to inhibit mitochondria.
  • Caffeine-induced calcium transients decayed 7- to 8-fold more slowly than electrically stimulated transients, indicating the sarcoplasmic reticulum accounts for 87% of total calcium removal.
  • Inhibition of Na-Ca exchange with Ni2+ decreased the caffeine response decay rate to approximately 33% of control.
  • The total relative contributions were estimated at 87% for the sarcoplasmic reticulum, 8.7% for Na-Ca exchange (accounting for 67% of non-sarcoplasmic reticulum removal), 2.6% for sarcolemmal Ca-ATPase, and 1.7% for mitochondria.

Structured PICO

P
Population
Isolated rat ventricular myocytes loaded with the fluorescent indicator indo-1
I
Intervention
Application of caffeine, Ni2+, FCCP, oligomycin, and varying extracellular calcium concentrations to isolate calcium removal mechanisms
C
Comparator
Control (electrically stimulated calcium transient or uninhibited caffeine response)
O
Outcome
Relative contributions of various intracellular and sarcolemmal systems to the relaxation of the systolic calcium transient (calcium removal)surrogate

In rat ventricular myocytes, the sarcoplasmic reticulum accounts for 87% of calcium removal during relaxation, while Na-Ca exchange accounts for 67% of the remaining non-SR mediated removal, highlighting a greater role for Ca-ATPase compared to rabbit models.

Abstract

OBJECTIVE: The aim was to estimate the relative contributions of the various intracellular and sarcolemmal systems to the relaxation of the systolic calcium transient. METHODS: The experiments were performed on isolated rat ventricular myocytes. The cells were loaded with the fluorescent indicator indo-1 in order to measure Ca2+i. RESULTS: The application of caffeine to release calcium from the sarcoplasmic reticulum produced a rise of Ca2+i which decayed about 7-8 times more slowly than the electrically stimulated calcium transient. This suggests that the sarcoplasmic reticulum accounts for about 87% of the calcium removal. The rate of decay of the caffeine response was decreased to about 33% of the control by inhibiting the Na-Ca exchange with Ni2+. In the presence of Ni2+ the rate could be inhibited further by inhibiting either the sarcolemmal Ca-ATPase (by increasing extracellular calcium concentration, Ca2+o) or the mitochondria (with FCCP and oligomycin). The relative contributions of the various processes were estimated to be: sarcoplasmic reticulum 87%, mitochondria 1.7%, Na-Ca 8.7%, sarcolemmal Ca-ATPase 2.6%. CONCLUSIONS: These experiments show that the Na-Ca exchange accounts for 67% of the calcium removal not mediated by the sarcoplasmic reticulum. This is a smaller fraction than in rabbit cardiac cells and highlights the importance of the Ca-ATPase in the rat heart.

Ask AI
Helpful
Bookmark
Share
View Full Paper

Cite This Study

Negretti et al. (1993) studied Isolated rat ventricular myocytes. Caffeine and various inhibitors (Ni2+, FCCP, oligomycin) vs. Control (electrically stimulated calcium transient) was evaluated on Relative contributions of intracellular and sarcolemmal systems to the relaxation of the systolic calcium transient. Calcium removal in isolated rat ventricular myocytes was primarily mediated by the sarcoplasmic reticulum (87%), followed by Na-Ca exchange (8.7%), sarcolemmal Ca-ATPase (2.6%), and mitochondria (1.7%).

synapsesocial.com/papers/6a15c5cc0c3a39952e9f9b47https://doi.org/10.1093/cvr/27.10.1826
Ask AI
Helpful
Bookmark
Share
View Full Paper