Real-time measurement of the length of a single sarcomere in rat ventricular myocytes: A novel analysis with quantum dots

Takahiro Serizawa, Takako Terui, Tatsuya Kagemoto, Akari Mizuno, Togo Shimozawa, Fuyu Kobirumaki, Shin'ichi Ishiwata, Satoshi Kurihara, Norio Fukuda*

*この研究の対応する著者

研究成果: Article査読

21 被引用数 (Scopus)

抄録

As the dynamic properties of cardiac sarcomeres are markedly changed in response to a length change of even ~0.1 μm, it is imperative to quantitatively measure sarcomere length (SL). Here we show a novel system using quantum dots (QDs) that enables a real-time measurement of the length of a single sarcomere in cardiomyocytes. First, QDs were conjugated with anti-α-actinin antibody and applied to the sarcomeric Z disks in isolated skinned cardiomyocytes of the rat. At partial activation, spontaneous sarcomeric oscillations (SPOC) occurred, and QDs provided a quantitative measurement of the length of a single sarcomere over the broad range (i.e., from ~1.7 to ~2.3 μm). It was found that the SPOC amplitude was inversely related to SL, but the period showed no correlation with SL. We then treated intact cardiomyocytes with the mixture of the antibody-QDs and FuGENE HD, and visualized the movement of the Z lines/T tubules. At a low frequency of 1 Hz, the cycle of the motion of a single sarcomere consisted of fast shortening followed by slow relengthening. However, an increase in stimulation frequency to 3-5 Hz caused a phase shift of shortening and relengthening due to acceleration of relengthening, and the waveform became similar to that observed during SPOC. Finally, the anti-α-actinin antibody-QDs were transfected from the surface of the beating heart in vivo. The striated patterns with ~1.96-μm intervals were observed after perfusion under fluorescence microscopy, and an electron microscopic observation confirmed the presence of QDs in and around the T tubules and Z disks, but primarily in the T tubules, within the first layer of cardiomyocytes of the left ventricular wall. Therefore, QDs are a useful tool to quantitatively analyze the movement of single sarcomeres in cardiomyocytes, under various experimental settings.

本文言語English
ジャーナルAmerican Journal of Physiology - Cell Physiology
301
5
DOI
出版ステータスPublished - 2011 11月
外部発表はい

ASJC Scopus subject areas

  • 細胞生物学
  • 生理学

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