Visualization of ATP with luciferin-luciferase reaction in mouse skeletal muscles using an "in vivo cryotechnique"

N. Terada, Y. Saitoh, S. Saitoh, N. Ohno, K. Fujishita, S. Koizumi, S. Ohno

研究成果: Article査読

4 被引用数 (Scopus)

抄録

Adenosine triphosphate (ATP) is a well-known energy source for muscle contraction. In this study, to visualize localization of ATP, a luciferin-luciferase reaction (LLR) was performed in mouse skeletal muscle with an "in vivo cryotechnique" (IVCT). First, to confirm if ATP molecules could be trapped and detected after glutaraldehyde (GA) treatment, ATP was directly attached to glass slides with GA, and LLR was performed. The LLR was clearly detected as an intentional design of the ATP attachment. The intensity of the light unit by LLR was correlated with the concentration of the GA-treated ATP in vitro. Next, LLR was evaluated in mouse skeletal muscles with IVCT followed by freeze-substitution fixation (FS) in acetone-containing GA. In such tissue sections the histological structure was well maintained, and the intensity of LLR in areas between muscle fibers and connective tissues was different. Moreover, differences in LLR among muscle fibers were also detected. For the IVCT-FS tissue sections, diaminobenzidine (DAB) reactions were clearly detected in type I muscle fibers and erythrocytes in capillaries, which demonstrated flow shape. Thus, it became possible to perform microscopic evaluation of the numbers of ATP molecules in the mouse skeletal muscles with IVCT, which mostly reflect living states.

本文言語English
ページ(範囲)1030-1036
ページ数7
ジャーナルMicroscopy and Microanalysis
18
5
DOI
出版ステータスPublished - 10-2012
外部発表はい

All Science Journal Classification (ASJC) codes

  • 器械工学

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