Lewis Katz School of Medicine at Temple University, Department of Neural Sciences, Shriners Pediatric Research Center, 3500 North Broad St, Philadelphia, PA 19140, USA.
J Cell Sci. 2023 Aug 15;136(16). doi: 10.1242/jcs.261356. Epub 2023 Aug 18.
The regulation of the intracellular level of ATP is a fundamental aspect of bioenergetics. Actin cytoskeletal dynamics have been reported to be an energetic drain in developing neurons and platelets. We addressed the role of actin dynamics in primary embryonic chicken neurons using luciferase assays, and by measurement of the ATP/ADP ratio using the ratiometric reporter PercevalHR and the ATP level using the ratiometric reporter mRuby-iATPSnFR. None of the methods revealed an effect of suppressing actin dynamics on the decline in the neuronal ATP level or the ATP/ADP ratio following shutdown of ATP production. Similarly, we find that treatments that elevate or suppress actin dynamics do not alter the ATP/ADP ratio in growth cones, the subcellular domain with the highest actin dynamics in developing neurons. Collectively, the data indicate that actin cytoskeletal dynamics are not a significant energy drain in developing neurons and that the ATP/ADP ratio is maintained when energy utilization varies. Discrepancies between prior work and the current data are discussed with emphasis on methodology and interpretation of the data.
细胞内 ATP 水平的调节是生物能量学的一个基本方面。已有报道称,在发育中的神经元和血小板中,肌动蛋白细胞骨架动力学是能量的消耗源。我们使用荧光素酶测定法和比率报告 PercevalHR 测量 ATP/ADP 比值以及比率报告 mRuby-iATPSnFR 测量 ATP 水平,在原代鸡胚神经元中研究了肌动蛋白动力学的作用。这些方法都没有揭示出抑制肌动蛋白动力学对 ATP 产生停止后神经元 ATP 水平或 ATP/ADP 比值下降的影响。同样,我们发现,升高或降低肌动蛋白动力学的处理方法不会改变生长锥中的 ATP/ADP 比值,生长锥是发育中的神经元中肌动蛋白动力学最高的亚细胞域。总的来说,这些数据表明,肌动蛋白细胞骨架动力学不是发育中的神经元的主要能量消耗源,并且当能量利用发生变化时,ATP/ADP 比值保持不变。当前数据与先前工作之间的差异在强调方法学和数据解释的基础上进行了讨论。