Imaging axon pathfinding in Xenopus in vivo

Area of Science:

• Neuroscience
• Developmental Biology
• Cell Biology

Background:

• Axon pathfinding is a dynamic process involving growth cone navigation and rapid cellular decisions.
• Visualizing axon pathfinding in vivo is essential for understanding growth cone responses to cues.
• Xenopus embryos offer advantages for live microscopy due to ex utero development and robustness.

Purpose of the Study:

• To describe a protocol for stabilizing and imaging live Xenopus embryos for high-resolution axon pathfinding visualization.
• To enable the study of gene function knockdown effects on axon navigation speed.
• To facilitate investigation of molecular mechanisms underlying growth cone behavior.

Main Methods:

• Protocol for stabilizing and preparing live Xenopus embryos for extended live microscopy (up to 72 hours).
• Utilizing Xenopus' accessible major axon tracts for imaging labeled axonal tracts.
• High spatial and temporal resolution imaging of axonal tracts in vivo.
• Combining live imaging with loss-of-function (e.g., gene knockdown) techniques.

Main Results:

• The protocol allows for prolonged, high-resolution imaging of axon pathfinding in live Xenopus embryos.
• Demonstrates the feasibility of investigating the impact of gene function knockdown on navigation speed.
• Successfully visualizes dynamic growth cone behaviors during navigation through complex territories.

Conclusions:

• This Xenopus-based live imaging protocol is a powerful tool for studying axon pathfinding dynamics.
• It facilitates the investigation of molecular cues and genetic factors influencing growth cone navigation.
• The adaptable method can be applied to various axon tracts and developmental processes.