The receptor phosphatase HmLAR2 collaborates with focal adhesion proteins in filopodial tips to control growth cone morphology

Dev Biol. 2008 Aug 1;320(1):215-25. doi: 10.1016/j.ydbio.2008.05.522. Epub 2008 May 17.

Abstract

Receptor protein tyrosine phosphatases (RPTPs) have been shown to play key roles in regulating axon guidance and synaptogenesis. HmLAR2, one of two closely related LAR-like RPTPs in the embryonic leech, is expressed in a few central neurons and in a unique segmentally-iterated peripheral cell, the comb cell (CC). Here we show that tagged HmLAR2-EGFP has a punctate pattern of expression in the growth cones of the CC, particularly at the tips of extending filopodia. Moreover, although expression of the wild-type EGFP-tagged receptor does not affect CC growth cone morphology, expression of a putative dominant-negative mutant of the receptor, CS-HmLAR2, leads to the enlargement of the growth cones, a shortening of filopodia, and errors in cellular tiling. RNAi of several candidate substrate signaling proteins, Lena (leech Ena/Vasp), beta-integrin and paxillin, but not beta-catenin, phenocopies particular aspects of the effects of HmLAR2 RNAi. For paxillin, which co-localizes with HmLAR2 at growth cone puncta, knock-down led to a reduction in the number of such puncta. Together, our data suggests that HmLAR2 regulates the morphology of the growth cone by controlling F-actin polymerization and focal adhesion complexes.

MeSH terms

  • Animals
  • CHO Cells
  • Catalysis
  • Cricetinae
  • Cricetulus
  • Focal Adhesions / enzymology*
  • Gene Expression Regulation, Developmental
  • Green Fluorescent Proteins / metabolism
  • Growth Cones / enzymology*
  • Leeches / cytology
  • Leeches / embryology
  • Leeches / enzymology*
  • Leeches / genetics
  • Mutation / genetics
  • Paxillin / metabolism
  • Phenotype
  • Phosphotyrosine / metabolism
  • Protein Binding
  • Protein Transport
  • Protein Tyrosine Phosphatases / metabolism*
  • Pseudopodia / enzymology*
  • RNA Interference
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction
  • Transgenes

Substances

  • Paxillin
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Phosphotyrosine
  • HmLAR2 protein, Hirudo medicinalis
  • Protein Tyrosine Phosphatases