WGA-Alexa Conjugates for Axonal Tracing

Curr Protoc Neurosci. 2017 Apr 10:79:1.28.1-1.28.24. doi: 10.1002/cpns.28.

Abstract

Anatomical labeling approaches are essential for understanding brain organization. Among these approaches are various methods of performing tract tracing. However, a major hurdle to overcome when marking neurons in vivo is visibility. Poor visibility makes it challenging to image a desired neuronal pathway so that it can be easily differentiated from a closely neighboring pathway. As a result, it becomes impossible to analyze individual projections or their connections. The tracer that is chosen for a given purpose has a major influence on the quality of the tracing. Here, we describe the wheat germ agglutinin (WGA) tracer conjugated to Alexa fluorophores for reliable high-resolution tracing of central nervous system projections. Using the mouse cerebellum as a model system, we implement WGA-Alexa tracing for marking and mapping neural circuits that control motor function. We also show its utility for marking localized regions of the cerebellum after performing single-unit extracellular recordings in vivo. © 2017 by John Wiley & Sons, Inc.

Keywords: WGA-Alexa; cerebellum; circuit tracing; connectivity; topography.

MeSH terms

  • Animals
  • Axons / metabolism*
  • Cerebellum / physiology*
  • Mice
  • Nerve Net / pathology
  • Neuroanatomical Tract-Tracing Techniques / methods*
  • Wheat Germ Agglutinins / metabolism*

Substances

  • Wheat Germ Agglutinins