Mechanical spinal cord transection in larval zebrafish and subsequent whole-mount histological processing

Zebrafish can regenerate their spinal cord after injury. Larval zebrafish are used to study spinal cord injury and regeneration because they are transparent, amenable to high-throughput analysis and have a rapid regeneration time. This protocol describes how to mechanically transect the spinal cord of larval zebrafish. It includes whole-mount tissue processing for in situ hybridization and immunohistochemistry. These techniques can elucidate the principles of regeneration.

• 1
  The protocol details two methods for spinal cord transection: incision and perforation lesion, both yielding comparable regeneration kinetics with reproducibility, high throughput, and low mortality.
• 2
  Axonal regrowth and functional recovery can be observed within two days after spinal cord transection in 3 dpf zebrafish.
• 3
  The described methods facilitate visualization of anatomical regeneration, interactions of regrowing neurites, and lesion-induced alterations in gene expression and protein localization.