Center for Ecology and Evolutionary Biology, Department of Biology, University of Oregon, Eugene OR 97403, USA.
Comp Biochem Physiol A Mol Integr Physiol. 2011 Sep;160(1):72-80. doi: 10.1016/j.cbpa.2011.05.005. Epub 2011 May 14.
Frog skeletal muscle mainly utilizes the substrates glucose and lactate for energy metabolism. The goal of this study was to determine the effect of insulin on the uptake and metabolic fate of lactate and glucose at rest in skeletal muscle of the American bullfrog, Lithobates catesbeiana, under varying temperature regimens. We hypothesize that lactate and glucose metabolic pathways will respond differently to the presence of insulin in cold versus warm acclimated frog tissues, suggesting an interaction between temperature and metabolism under varying environmental conditions. We employed radiolabeled tracer techniques to measure in vitro uptake, oxidation, and incorporation of glucose and lactate into glycogen by isolated muscles from bullfrogs acclimated to 5 °C (cold) or 25 °C (warm). Isolated bundles from Sartorius muscles were incubated at 5 °C, 15 °C, or 25 °C, and in the presence and absence of 0.05 IU/mL bovine insulin. Insulin treatment in the warm acclimated and incubated frogs resulted in an increase in glucose incorporation into glycogen, and an increase in intracellular [glucose] of 0.5 μmol/g (P<0.05). Under the same conditions lactate incorporation into glycogen was reduced (P<0.05) in insulin-treated muscle. When compared to the warm treatment group, cold acclimation and incubation resulted in increased rates of glucose oxidation and glycogen synthesis, and a reduction in free intracellular glucose levels (P<0.05). When muscles from either acclimation group were incubated at an intermediate temperature of 15 °C, insulin's effect on substrate metabolism was attenuated or even reversed. Therefore, a significant interaction between insulin and acclimation condition in controlling skeletal muscle metabolism appears to exist. Our findings further suggest that one of insulin's actions in frog muscle is to increase glucose incorporation into glycogen, and to reduce reliance on lactate as the primary metabolic fuel.
蛙类骨骼肌主要利用葡萄糖和乳酸作为能量代谢底物。本研究旨在确定胰岛素对不同温度条件下,美牛蛙(Lithobates catesbeiana)骨骼肌内乳酸和葡萄糖摄取及其代谢命运的影响。我们假设在冷适应和暖适应蛙组织中,乳酸和葡萄糖代谢途径对胰岛素的存在会有不同的反应,这表明在不同环境条件下,温度和代谢之间存在相互作用。我们采用放射性示踪技术测量了来自适应于 5°C(冷)或 25°C(暖)的牛蛙的离体肌肉对葡萄糖和乳酸的体外摄取、氧化和掺入糖原的情况。在 5°C、15°C 或 25°C 下孵育来自 Sartorius 肌肉的分离束,并在存在和不存在 0.05 IU/mL 牛胰岛素的情况下进行孵育。在暖适应和孵育的青蛙中,胰岛素处理导致葡萄糖掺入糖原增加,细胞内[葡萄糖]增加 0.5 μmol/g(P<0.05)。在胰岛素处理的肌肉中,乳酸掺入糖原减少(P<0.05)。与暖处理组相比,冷适应和孵育导致葡萄糖氧化和糖原合成速率增加,细胞内游离葡萄糖水平降低(P<0.05)。当来自任一适应组的肌肉在中间温度 15°C 下孵育时,胰岛素对底物代谢的影响减弱甚至逆转。因此,胰岛素和适应条件之间在控制骨骼肌代谢方面似乎存在显著的相互作用。我们的研究结果进一步表明,胰岛素在蛙类肌肉中的作用之一是增加葡萄糖掺入糖原,并减少对乳酸作为主要代谢燃料的依赖。