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Simultaneous imaging of local calcium and single sarcomere length in rat neonatal cardiomyocytes using yellow Cameleon-Nano140

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ABSTRACT

Excitation–contraction coupling results in the shortening of many individual sarcomeres along the length of a muscle fiber. Tsukamoto and colleagues develop a technique to quantitatively analyze the dynamics of intracellular calcium transients and length changes at the single sarcomere level.

No MeSH data available.


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Comparison of a time course of CaT detected by α-actinin–YC-Nano140 with that by Fluo-4 in cardiomyocytes at different temperatures during spontaneous beating. (A) Time course of a change in Fyellow/Fcyan (α-actinin–YC-Nano140) or Fluo-4 fluorescence in cardiomyocytes at 22°C and 37°C. CaT was normalized at the peak for all curves. Each curve was averaged from five beats obtained in 8–24 cells (see B for experimental numbers). (B) Graph comparing the time to peak values for α-actinin–YC-Nano140 or Fluo-4 at 22°C and 37°C. n = 57 beats (8 cells), 61 beats (8 cells), 514 beats (24 cells), and 203 beats (14 cells) for Fluo-4 (22°C), α-actinin–YC-Nano140 (22°C), Fluo-4 (37°C), and α-actinin–YC-Nano140 (37°C), respectively. (C) Graph comparing the decay time values for α-actinin–YC-Nano140 and Fluo-4 at 22°C and 37°C. n, same as in B. (B and C) ***, P < 0.001 (Tukey-Kramer test). Bars in each graph indicate mean values.
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fig3: Comparison of a time course of CaT detected by α-actinin–YC-Nano140 with that by Fluo-4 in cardiomyocytes at different temperatures during spontaneous beating. (A) Time course of a change in Fyellow/Fcyan (α-actinin–YC-Nano140) or Fluo-4 fluorescence in cardiomyocytes at 22°C and 37°C. CaT was normalized at the peak for all curves. Each curve was averaged from five beats obtained in 8–24 cells (see B for experimental numbers). (B) Graph comparing the time to peak values for α-actinin–YC-Nano140 or Fluo-4 at 22°C and 37°C. n = 57 beats (8 cells), 61 beats (8 cells), 514 beats (24 cells), and 203 beats (14 cells) for Fluo-4 (22°C), α-actinin–YC-Nano140 (22°C), Fluo-4 (37°C), and α-actinin–YC-Nano140 (37°C), respectively. (C) Graph comparing the decay time values for α-actinin–YC-Nano140 and Fluo-4 at 22°C and 37°C. n, same as in B. (B and C) ***, P < 0.001 (Tukey-Kramer test). Bars in each graph indicate mean values.

Mentions: Next, we investigated the rate of rise or fall of the fluorescence signal of α-actinin–YC-Nano140, in comparison with Fluo-4, a commonly used small molecule Ca2+ indicator (compare Shintani et al., 2014), as a reference. We found that the time course of the Fyellow/Fcyan duration of α-actinin–YC-Nano140 was longer than that of Fluo-4, at both low (i.e., 22°C; ∼2 and ∼1 s for α-actinin–YC-Nano140 and Fluo-4, respectively) and high (i.e., 37°C; ∼1.2 and ∼0.7 s for α-actinin–YC-Nano140 and Fluo-4, respectively) temperatures (Fig. 3 A). In addition, time to peak and decay time were both significantly longer for α-actinin–YC-Nano140 than Fluo-4 at both temperatures (Fig. 3, B and C).


Simultaneous imaging of local calcium and single sarcomere length in rat neonatal cardiomyocytes using yellow Cameleon-Nano140
Comparison of a time course of CaT detected by α-actinin–YC-Nano140 with that by Fluo-4 in cardiomyocytes at different temperatures during spontaneous beating. (A) Time course of a change in Fyellow/Fcyan (α-actinin–YC-Nano140) or Fluo-4 fluorescence in cardiomyocytes at 22°C and 37°C. CaT was normalized at the peak for all curves. Each curve was averaged from five beats obtained in 8–24 cells (see B for experimental numbers). (B) Graph comparing the time to peak values for α-actinin–YC-Nano140 or Fluo-4 at 22°C and 37°C. n = 57 beats (8 cells), 61 beats (8 cells), 514 beats (24 cells), and 203 beats (14 cells) for Fluo-4 (22°C), α-actinin–YC-Nano140 (22°C), Fluo-4 (37°C), and α-actinin–YC-Nano140 (37°C), respectively. (C) Graph comparing the decay time values for α-actinin–YC-Nano140 and Fluo-4 at 22°C and 37°C. n, same as in B. (B and C) ***, P < 0.001 (Tukey-Kramer test). Bars in each graph indicate mean values.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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fig3: Comparison of a time course of CaT detected by α-actinin–YC-Nano140 with that by Fluo-4 in cardiomyocytes at different temperatures during spontaneous beating. (A) Time course of a change in Fyellow/Fcyan (α-actinin–YC-Nano140) or Fluo-4 fluorescence in cardiomyocytes at 22°C and 37°C. CaT was normalized at the peak for all curves. Each curve was averaged from five beats obtained in 8–24 cells (see B for experimental numbers). (B) Graph comparing the time to peak values for α-actinin–YC-Nano140 or Fluo-4 at 22°C and 37°C. n = 57 beats (8 cells), 61 beats (8 cells), 514 beats (24 cells), and 203 beats (14 cells) for Fluo-4 (22°C), α-actinin–YC-Nano140 (22°C), Fluo-4 (37°C), and α-actinin–YC-Nano140 (37°C), respectively. (C) Graph comparing the decay time values for α-actinin–YC-Nano140 and Fluo-4 at 22°C and 37°C. n, same as in B. (B and C) ***, P < 0.001 (Tukey-Kramer test). Bars in each graph indicate mean values.
Mentions: Next, we investigated the rate of rise or fall of the fluorescence signal of α-actinin–YC-Nano140, in comparison with Fluo-4, a commonly used small molecule Ca2+ indicator (compare Shintani et al., 2014), as a reference. We found that the time course of the Fyellow/Fcyan duration of α-actinin–YC-Nano140 was longer than that of Fluo-4, at both low (i.e., 22°C; ∼2 and ∼1 s for α-actinin–YC-Nano140 and Fluo-4, respectively) and high (i.e., 37°C; ∼1.2 and ∼0.7 s for α-actinin–YC-Nano140 and Fluo-4, respectively) temperatures (Fig. 3 A). In addition, time to peak and decay time were both significantly longer for α-actinin–YC-Nano140 than Fluo-4 at both temperatures (Fig. 3, B and C).

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Excitation&ndash;contraction coupling results in the shortening of many individual sarcomeres along the length of a muscle fiber. Tsukamoto and colleagues develop a technique to quantitatively analyze the dynamics of intracellular calcium transients and length changes at the single sarcomere level.

No MeSH data available.


Related in: MedlinePlus