3D reconstruction of neuronal allometry and neuromuscular projections in asexual planarians using expansion tiling light sheet microscopy

• DOI: 10.7554/elife.101103.3
• Publication Date: 2025-03-28T20:13:37Z
• Authors: Jing Lu; Hao Xu; Dongyue Wang; Yanlu Chen; Takeshi Inoue; Liang Gao; Kai Lei
• Keywords: Allometry

The intricate coordination of the neural network in planarian growth and regeneration has remained largely unrevealed, partly due to challenges imaging CNS three dimensions (3D) with high resolution within a reasonable timeframe. To address this gap systematic planarians, we adopted high-resolution, nanoscale by combining tissue expansion tiling light-sheet microscopy, achieving up fourfold linear expansion. Using an automatic 3D cell segmentation pipeline, quantitatively profiled neurons muscle fibers at single-cell level over 400 wild-type planarians during homeostasis regeneration. We validated previous observations neuronal number changes fiber distribution. found that increase neuron tends lag behind rapid somatic cells later phase homeostasis. By 120 nm resolution, also observed distinct distribution patterns anterior posterior poles. Furthermore, investigated effects β-catenin-1 RNAi on pole, consistent anterior-posterior polarity. glial were be close contact dorsal-ventral fibers. Finally, disruptions neural-muscular networks inr-1 planarians. These findings provide insights into detailed structure potential functions system highlight accessibility our tool unraveling biological underlying their diverse phenotypes behaviors.