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?Fig.2.2. The excitation-induced net loss of K+ was Selleckchem BIBF-1120 calculated as the difference between total K+ contents of resting muscles and that of stimulated ... Table 2 Stimulation induced 22Na uptake and net increase in intracellular Na+ content in rat EDL muscles in vitro. All values for Na+ represent the increase over and above the 22Na uptake or Na+ content in resting muscles. The uptake per stimulation pulse is ... [K+]o was calculated by dividing the excitation-induced net loss of K+ by the 14C-sucrose space measured in EDL muscles in vitro (0.214?mL/g wet wt., Clausen et?al. 2004). After 15 and 60?sec of stimulation [K+]o increases by 44?mmol?L?1 (9.4?��mol/0.214?mL; P?GS-7340 mw (Clausen et?al. 2004). The mechanisms of the clearance of K+ from the extracellular space was previously (Clausen 2008a) examined by comparing the loss of K+ during stimulation in KR buffer with muscles where the buffer had been removed, preventing the clearance by diffusion into surrounding buffer. After 30?sec of 60?Hz stimulation, the average concentration of K+ in the interstitial water space reached the same level (48.6?mmol?L?1) as in the muscles stimulated in air (46.7?mmol?L?1) (N?=?9 vs. 7, P?=?0.87). This indicates that K+ is not primarily cleared by diffusion, but by reaccumulation via the Na+,K+ pumps into the cells. In keeping with this, the omission of KR buffer caused no interference Adenylyl cyclase with contractile performance, when compared with muscles kept in KR buffer neither in EDL, nor in soleus muscle (Clausen 2008a). In contrast, ouabain-induced inhibition of K+ clearance via the Na+,K+ pumps markedly reduced contractile endurance and force recovery in air (Clausen 2008a). In keeping with this, ouabain as well as downregulation of the muscle contents of Na+,K+ pumps induced by prior K+ depletion of the rats were found to induce a graded reduction of the contractile endurance in rat soleus (Nielsen and Clausen 1996). The effects of increasing the incubation temperature from 30 to 37��C on excitation-induced 22Na uptake was examined. As shown in Figure?Figure4A4A and B as compared with Figure?Figure2A2A and B, respectively, this increase in temperature caused no significant change in the effect of 60?Hz stimulation on 22Na uptake or Na+ content.