The metabolic turnover in the isolated in vitro perfused and superfused rat skeletal muscle (musculus gracilis cranialis) was enhanced by increasing the medium flow rate under relaxed conditions. In a recent study we have measured the tissue concentrations of second messengers: cyclic adenosine 3 ́5 ́- monophosphate (cAMP), cyclic guanosine 3 ́5 ́-
monophosphate (cGMP), and D-myo-inositol 1, 4, 5-trisphosphate (IP3) under similar experimental conditions to analyze their potential role in the described stimulation of metabolic rate by changes of perfusion flow rate. The tissue levels of the two second messengers’ cAMP and cGMP were not significantly changed after increasing the perfusion
flow rate and they probably have no transduction role in the induced alteration of skeletal muscle metabolism. However, the IP3 content was extremely reduced after increasing flow rate. This decrease in the tissue concentration of IP3 induced by increasing the flow rate indicates the possible role of IP3 in this signal transduction, leading to changes in
the cellular metabolic pathways.
Adenosine 5'-triphosphate (ATP), phosphocreatine (PCr), creatine (Cr), inorganic phosphate (Pi), lactate (LAC), pyruvate (PYR) and glycogen as glucose (GLG) were determined and free adenosine 5'-diphosphate (ADP) was calculated from ATP:creatine phosphokinase (CPK) reaction in the gracilis muscle of cold-acclimated rats in vivo, and in completely isolated muscles under medium perfusion and superfusion in vitro, using the freezeclamping method. The mean in vivo levels (wmol/g w.w.) were: ATP 4.8, PCr 12.0, Cr 7.8, Pi 16.1, LAC 1.6, PYR 0.09, GLG 22.9, ADP 0.62 x 10~3. Isolation of the muscle (about 11 min of anoxia followed by perfusion in the air with a high pC>2 medium) decreased macroergic phosphate levels (ATP 3.0 , PCr 8.3). In isolated muscles perfused with a high pC>2 medium (99 kPa O2, perfusion rate 70 /rl/min) and simultaneously superfused with a low pC>2 medium (6.2 kPa O2, 2.3 ml/min) at 28 °C in vitro the levels of metabolites were (wmol/g w.w.): ATP 3.1, PCr 8.5, Cr 5.6, Pi 9.2 LAC 2.1, PYR 0.19, GLG 6.6, ADP 0.44 x 10-3. The mean steady oxygen uptake of the isolated muscle was 97 nmol O2 x min-1 x g-1 w.w. Thus, the levels of macroergic phosphates and their derivatives are lower after isolation and perfusion of the muscle, but the creatine charge [PCr]/([PCr]-l-[Cr]] remains stable (0.61 in vivo versus 0.60 in the isolated muscle). This indicates that the steady-state and high energy status of the isolated perfused-superfused gracilis muscle is maintained.
Adenosine 5'-triphosphate (ATP), phosphocreatine (PCr), creatine (Cr), inorganic phosphate (Pi), lactate (LAC), pyruvate (PYR) and glycogen as glucose (GLG) were determined and free adenosine 5'-diphosphate (ADP) was calculated from ATP:creatine phosphokinase (CPK) reaction in the gracilis muscle of cold-acclimated rats in vivo, and in completely isolated muscles under medium perfusion and superfusion in vitro, using the freezeclamping method. The mean in vivo levels (wmol/g w.w.) were: ATP 4.8, PCr 12.0, Cr 7.8, Pi 1.6, LAC 1.6, PYR 0.09, GLG 22.9, ADP 0.62 x 10-3. Isolation of the muscle (about 11 min of anoxia followed by perfusion in the air with a high pC>2 medium) decreased macroergic phosphate levels (ATP 3.0 , PCr 8.3). In isolated muscles perfused with a high pC>2 medium (99 kPa O2, perfusion rate 70 /rl/min) and simultaneously superfused with a low p02 medium (6.2 kPa O2, 2.3 ml/min) at 28 °C in vitro the levels of metabolites were (wmol/g w.w.): ATP 3.1, PCr 8.5, Cr 5.6, Pi 0.9, LAC 2.1, PYR 0.19, GLG 6.6, ADP 0.44 x 10-3. The mean steady oxygen uptake of the isolated muscle was 97 nmol O2 x min-1 x g-1 w.w. Thus, the levels of macroergic phosphates and their derivatives are lower after isolation and perfusion of the muscle, but the creatine charge [PCr]/([PCr]-f[Cr]] remains stable (0.61 in vivo versus 0.60 in the isolated muscle). This indicates that the steady-state and high energy status of the isolated perfused-superfused gracilis muscle is maintained.
Fever developing after intracerebral injections of lipopolysaccharide (LPS) to guinea-pigs were monophasic, with only one peak of inner body temperature,slowly developing and longlasting in a dose range 20 to 200 ng of LPS. Latency time was inversely related to the dose of LPS. Indomethacin injected to the third brain ventricle did not abolish fever response.