Research data supporting: 'Auxin-inducible degradation of UNC-116 in <i>C. elegans</i> inhibits bidirectional dense core vesicle transport and worm locomotion on different timescales’

The microtubule motor kinesin-1 is vital in neurons, with mutations being associated with neurological diseases. Deletion of the <i>Caenorhabditis elegans</i> kinesin-1 gene <i>unc-116</i> is lethal, and viable mutants are uncoordinated. Here, we perform auxin-mediated degradation of the UNC-116 protein and monitor effects on worm locomotion and dense core vesicle (DCV) transport. UNC-116 is degraded within 1-3 hours of auxin treatment. Impaired swimming/thrashing and crawling are detected after 6-14 hours, depending on organismal age, and degradation of UNC-116 in neurons alone disrupts locomotion. Effects on DCV transport are observed sooner, with 4 hours of degradation strongly inhibiting both anterograde and retrograde movement. This motility recovers when worms are transferred to auxin-free plates for 24 hours, even though protein level recovery is incomplete. Rescue from a 10-hour auxin treatment completely restores crawling speed, but although swimming is initially unaffected, it becomes strongly inhibited during the rescue period, suggesting that UNC-116 has distinct roles in these locomotive gaits. Overall, by bypassing early developmental UNC-116 functions, we reveal that UNC-116 is essential for bidirectional DCV transport and crucial for worm locomotion.微管马达蛋白驱动蛋白-1（kinesin-1）在神经元中发挥关键作用，其突变与多种神经系统疾病密切相关。秀丽隐杆线虫（Caenorhabditis elegans）的驱动蛋白-1基因unc-116纯合缺失会导致个体死亡，而存活的突变体则表现出运动协调障碍。本研究通过生长素介导的途径降解UNC-116蛋白，并监测其对秀丽隐杆线虫运动以及致密核心囊泡（dense core vesicle，DCV）运输的影响。 生长素处理后1至3小时内，UNC-116即可被降解。根据线虫个体发育阶段的不同，在处理后6至14小时可观察到其游泳/摆躯运动以及爬行能力受损；仅在神经元中降解UNC-116即可破坏线虫的运动能力。而对致密核心囊泡运输的影响出现更早，仅降解4小时即可显著抑制顺向运输与逆向运输过程。 将线虫转移至不含生长素的培养平板上培养24小时后，其运动能力可恢复，即便此时UNC-116的蛋白水平尚未完全恢复。对经10小时生长素处理的线虫进行挽救培养，可完全恢复其爬行速度；尽管游泳运动最初未受影响，但在挽救培养期间会出现显著的运动抑制，这表明UNC-116在这两种运动步态中发挥着截然不同的功能。 综上，本研究通过规避UNC-116在发育早期的功能，揭示了其对于致密核心囊泡双向运输以及线虫运动能力均至关重要。