Osuma Edie A, Riggs Daniel W, Gibb Andrew A, Hill Bradford G
Wesleyan College Macon GA USA.
Department of Physiology University of Louisville Louisville KY USA.
Regeneration (Oxf). 2018 Jan 11;5(1):78-86. doi: 10.1002/reg2.95. eCollection 2018 Mar.
Planarians are outstanding models for studying mechanisms of regeneration; however, there are few methods to measure changes in their metabolism. Examining metabolism in planarians is important because the regenerative process is dependent on numerous integrated metabolic pathways, which provide the energy required for tissue repair as well as the ability to synthesize the cellular building blocks needed to form new tissue. Therefore, we standardized an extracellular flux analysis method to measure mitochondrial and glycolytic activity in live planarians during normal growth as well as during regeneration. Small, uninjured planarians showed higher rates of oxygen consumption compared with large planarians, with no difference in glycolytic activity; however, glycolysis increased during planarian regeneration. Exposure of planarians to koningic acid, a specific inhibitor of glyceraldehyde-3-phosphate dehydrogenase, completely abolished extracellular acidification with little effect on oxygen consumption, which suggests that the majority of glucose catabolized in planarians is fated for aerobic glycolysis. These studies describe a useful method for measuring respiration and glycolysis in planarians and provide data implicating changes in glucose metabolism in the regenerative response.
涡虫是研究再生机制的优秀模型;然而,测量其新陈代谢变化的方法却很少。研究涡虫的新陈代谢很重要,因为再生过程依赖于众多整合的代谢途径,这些途径为组织修复提供所需能量,以及合成形成新组织所需细胞构件的能力。因此,我们标准化了一种细胞外通量分析方法,以测量正常生长以及再生过程中活体涡虫的线粒体和糖酵解活性。与大型涡虫相比,小型、未受伤的涡虫显示出更高的耗氧率,糖酵解活性没有差异;然而,涡虫再生期间糖酵解增加。将涡虫暴露于3-磷酸甘油醛脱氢酶的特异性抑制剂米曲霉酸中,完全消除了细胞外酸化对耗氧的影响很小,这表明涡虫中分解代谢的大部分葡萄糖用于有氧糖酵解。这些研究描述了一种测量涡虫呼吸和糖酵解的有用方法,并提供了暗示再生反应中葡萄糖代谢变化的数据。