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A luminometric method for the determination of ATP and phosphocreatine in single human skeletal muscle fibres

✍ Scribed by Wibom, R. ;Söderlund, K. ;Lundin, A. ;Hultman, E.

Publisher
John Wiley and Sons
Year
1991
Weight
529 KB
Volume
6
Category
Article
ISSN
0884-3996

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✦ Synopsis

A sensitive method for the analysis o f ATP and phosphocreatine (PCr) in single human skeletal muscle fibres is described. Muscle tissue was freeze-dried and single fibres were dissected free w i t h the aid of low-power microscopy. The fibres were then extracted in trichloroacetic acid and neutralized with KHCO,. The assay is based o n the continuous monitoring o f light produced as a result o f ATP degradation in the firefly luciferase reaction. PCr is measured as the amount o f ATP formed in the creatine kinase reaction. The coefficient o f variation was less than 4 % for both ATP and PCr determination. The amount o f tissue required for the assay is approximately 0 . 5 ~~ (dry weight). The assay showed good agreement w i t h spectrophotometric and highperformance liquid chromatographic (HPLC) measurements made upon extracts o f whole muscle tissue.

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