Research article

Transient axonal glycoprotein-1 (TAG-1) and laminin-α1 regulate dynamic growth cone behaviors and initial axon direction in vivo

Marc A Wolman^1,4 , Vinoth K Sittaramane³ , Jeffrey J Essner^2,5 , H Joseph Yost² , Anand Chandrasekhar³ and Mary C Halloran¹

¹Departments of Zoology and Anatomy, and Neuroscience Training Program, University of Wisconsin, Madison, Wisconsin 53706, USA

²Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City, Utah 84112-5330, USA

³Division of Biological Sciences, University of Missouri, Columbia, Missouri 65211, USA

⁴Department of Cell and Developmental Biology, University of Pennsylvania Medical School, Philadelphia, PA 19104, USA

⁵Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA

author email corresponding author email

Neural Development 2008, 3:6doi:10.1186/1749-8104-3-6

Published:	20 February 2008

Abstract

Background

How axon guidance signals regulate growth cone behavior and guidance decisions in the complex in vivo environment of the central nervous system is not well understood. We have taken advantage of the unique features of the zebrafish embryo to visualize dynamic growth cone behaviors and analyze guidance mechanisms of axons emerging from a central brain nucleus in vivo.

Results

We investigated axons of the nucleus of the medial longitudinal fascicle (nucMLF), which are the first axons to extend in the zebrafish midbrain. Using in vivo time-lapse imaging, we show that both positive axon-axon interactions and guidance by surrounding tissue control initial nucMLF axon guidance. We further show that two guidance molecules, transient axonal glycoprotein-1 (TAG-1) and laminin-α1, are essential for the initial directional extension of nucMLF axons and their subsequent convergence into a tight fascicle. Fixed tissue analysis shows that TAG-1 knockdown causes errors in nucMLF axon pathfinding similar to those seen in a laminin-α1 mutant. However, in vivo time-lapse imaging reveals that while some defects in dynamic growth cone behavior are similar, there are also defects unique to the loss of each gene. Loss of either TAG-1 or laminin-α1 causes nucMLF axons to extend into surrounding tissue in incorrect directions and reduces axonal growth rate, resulting in stunted nucMLF axons that fail to extend beyond the hindbrain. However, defects in axon-axon interactions were found only after TAG-1 knockdown, while defects in initial nucMLF axon polarity and excessive branching of nucMLF axons occurred only in laminin-α1 mutants.

Conclusion

These results demonstrate how two guidance cues, TAG-1 and laminin-α1, influence the behavior of growth cones during axon pathfinding in vivo. Our data suggest that TAG-1 functions to allow growth cones to sense environmental cues and mediates positive axon-axon interactions. Laminin-α1 does not regulate axon-axon interactions, but does influence neuronal polarity and directional guidance.