Department of Ocean Sciences, Ocean Sciences Centre, Memorial University of Newfoundland, St John's, NL, Canada, A1C 5S7.
J Exp Biol. 2013 Feb 1;216(Pt 3):437-46. doi: 10.1242/jeb.079178. Epub 2012 Oct 4.
The aim of the present study was to assess whether mechanisms of glucose trafficking by red blood cells (RBCs) relate to species-specific extracellular glucose levels. Atlantic cod (Gadus morhua), Atlantic salmon (Salmo salar), cunner (Tautogolabrus adspersus) and short-horned sculpin (Myoxocephalus scorpius) had plasma glucose levels of 4, 4.1, 1.95 and 0.73 mmol l(-1), respectively. Glucose uptake by isolated RBCs was measured by the initial incorporation of [6-(14)C]-glucose and steady-state glucose metabolism was determined by the production of (3)H(2)O from [2-(3)H]-glucose. Saturation kinetics of glucose uptake and inhibition of both glucose uptake and metabolism by cytochalasin B and phloretin revealed that Atlantic cod, cunner and sculpin RBCs all had a facilitated transport component to glucose trafficking. RBCs from Atlantic salmon showed a linear relationship between glucose uptake and extracellular glucose level, but exhibited clear inhibition of glucose metabolism by cytochalasin B and phloretin, suggesting a component of facilitated glucose transport that is more elusive to detect. The production of (3)H(2)O was linear for at least 6 h and as such presents a rigorous approach to measuring glycolytic rate. Steady-state rates of glucose metabolism were achieved at extracellular levels of approximately 1 mmol l(-1) glucose for RBCs from all species, showing that within-species normal extracellular glucose level is not a primary determinant of the basal level of glycolysis. At physiological levels of extracellular glucose, the ratio of initial glucose uptake to glucose metabolism was 1.5 to 4 for all RBCs, suggesting that there is scope to increase metabolic rate without alteration of the basal glucose uptake capacity.
本研究旨在评估红细胞(RBC)摄取葡萄糖的机制是否与物种特异性的细胞外葡萄糖水平有关。大西洋鳕鱼(Gadus morhua)、大西洋三文鱼(Salmo salar)、圆鳍鱼(Tautogolabrus adspersus)和短须石首鱼(Myoxocephalus scorpius)的血浆葡萄糖水平分别为 4、4.1、1.95 和 0.73mmol·l(-1)。通过初始掺入[6-(14)C]-葡萄糖测量分离的 RBC 对葡萄糖的摄取,通过[2-(3)H]-葡萄糖产生(3)H2O 来确定葡萄糖代谢的稳态。葡萄糖摄取的饱和动力学以及细胞松弛素 B 和根皮苷对葡萄糖摄取和代谢的抑制作用表明,大西洋鳕鱼、圆鳍鱼和短须石首鱼的 RBC 均具有促进葡萄糖转运的组成部分。大西洋三文鱼的 RBC 摄取葡萄糖与细胞外葡萄糖水平之间呈线性关系,但细胞松弛素 B 和根皮苷对葡萄糖代谢的抑制作用明显,表明存在更难以检测到的促进葡萄糖转运组成部分。(3)H2O 的产生至少在 6 小时内呈线性,因此是测量糖酵解率的严格方法。所有物种的 RBC 在细胞外葡萄糖水平约为 1mmol·l(-1)时达到葡萄糖代谢的稳态速率,表明同种内正常的细胞外葡萄糖水平不是糖酵解基础水平的主要决定因素。在细胞外葡萄糖的生理水平下,所有 RBC 的初始葡萄糖摄取与葡萄糖代谢的比值为 1.5 至 4,表明在不改变基础葡萄糖摄取能力的情况下,代谢率仍有提高的空间。