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  • The CSA measured at a defined sarcomere length in individual single muscle fibres decreased progressively in response to NMB and muscle unloading at durations of 5 days and longer, irrespective of MyHC isoform expression according to one-way ANOVA (P < 0.001, Fig. 2A). In contrast to enzyme-histochemistry, muscle fibres could be separated into specific types based on MyHC isoform expression at all time-points due to the higher sensitivity of the silver-stained MK 2206 SDS-PAGE, in accordance with previous observations in ICU patients with AQM (Larsson et al. 2000; Norman et al. 2009). Maximum force normalized to muscle fibre cross-sectional area (specific force) and stiffness did not differ between soleus fibres expressing the type I MyHC isoform or EDL fibres expressing the IIx or IIb MyHC isoform at any of the different durations and have therefore been pooled. Both specific force and stiffness were similar in the control and 0.25�C4 day groups, but decreased significantly at longer NMB durations (Fig. 2B and C). Stiffness is dependent on the number of attached cross-bridges, force/compliance of each cross-bridge and compliance of the structures Selleck R428 in series with the cross-bridges (Higuchi et al. 1995, Seow et al. 1997, Galler & Hilber, 1998). The strong linear relationship between stiffness and force irrespective of whether fibres were from controls or from rats exposed to 2 weeks of NMB (not shown) underscores that stiffness is primarily dependent on the number of attached cross-bridges (Regnier et al. 2004). The subsequent specific force:stiffness ratio may, therefore, be closely related to the force/compliance of each cross-bridge (McDonald & Fitts, 1995). The unchanged specific force:stiffness ratios for all the groups, suggest that the force/compliance of each cross-bridge was preserved and the decreased specific force was primarily due to a reduced number of force-generating cross-bridges (Fig. 2D). This is consistent with the biochemical analyses demonstrating a preferential loss of the motor protein Epigenetics inhibitor myosin at durations longer than 4 days (see below). A strong linear relationship was also observed between specific force and the myosin:actin ratio in muscle fibres expressing the type IIb (r2= 0.81; P < 0.001) and IIx (r2= 0.40; P < 0.01) MyHC isoforms. In soleus muscle fibres expressing the type I MyHC isoform, on the other hand, this relationship was weak and not statistically significant. The exact mechanism underlying the muscle fibre type-specific differences in this relationship is not known, but may be related to fibre type-specific differences in total amount of contractile proteins in individual muscle fibres or in intrinsic force-generating properties of specific myosin isoforms.

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