Flotillin-mediated endocytic events dictate cell type-specific responses to semaphorin 3A

J Neurosci. 2010 Nov 10;30(45):15317-29. doi: 10.1523/JNEUROSCI.1821-10.2010.

Abstract

Cortical efferents growing in the same environment diverge early in development. The expression of particular transcription factors dictates the trajectories taken, presumably by regulating responsiveness to guidance cues via cellular mechanisms that are not yet known. Here, we show that cortical neurons that are dissociated and grown in culture maintain their cell type-specific identities defined by the expression of transcription factors. Using this model system, we sought to identify and characterize mechanisms that are recruited to produce cell type-specific responses to Semaphorin 3A (Sema3A), a guidance cue that would be presented similarly to cortical axons in vivo. Axons from presumptive corticofugal neurons lacking the transcription factor Satb2 and expressing Ctip2 or Tbr1 respond far more robustly to Sema3A than those from presumptive callosal neurons expressing Satb2. Both populations of axons express similar levels of Sema3A receptors (neuropilin-1, cell adhesion molecule L1, and plexinA4), but significantly, axons from neurons lacking Satb2 internalize more Sema3A, and they do so via a raft-mediated endocytic pathway. We used an in silico approach to identify the endocytosis effector flotillin-1 as a Sema3A signaling candidate. We tested the contributions of flotillin-1 to Sema3A endocytosis and signaling, and show that raft-mediated Sema3A endocytosis is defined by and depends on the recruitment of flotillin-1, which mediates LIM domain kinase activation and regulates axon responsiveness to Sema3A in presumptive corticofugal axons.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Analysis of Variance
  • Animals
  • Axons / drug effects
  • Axons / metabolism*
  • Blotting, Western
  • Cerebral Cortex / cytology
  • Cerebral Cortex / drug effects
  • Cerebral Cortex / metabolism
  • Endocytosis / drug effects
  • Endocytosis / physiology*
  • Matrix Attachment Region Binding Proteins / genetics
  • Matrix Attachment Region Binding Proteins / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Neural Cell Adhesion Molecule L1 / genetics
  • Neural Cell Adhesion Molecule L1 / metabolism
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism*
  • RNA Interference
  • Rats
  • Rats, Sprague-Dawley
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Semaphorin-3A / metabolism*
  • Semaphorin-3A / pharmacology
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • T-Box Domain Proteins / genetics
  • T-Box Domain Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism

Substances

  • Bcl11b protein, mouse
  • Matrix Attachment Region Binding Proteins
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Neural Cell Adhesion Molecule L1
  • Repressor Proteins
  • SATB2 protein, mouse
  • Semaphorin-3A
  • T-Box Domain Proteins
  • Tbr-1 protein, rat
  • Transcription Factors
  • Tumor Suppressor Proteins
  • flotillins