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Analysis of Schwann-astrocyte Interactions Using In Vitro Assays
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Direct Thy-1/alphaVbeta3 integrin interaction mediates neuron to astrocyte communication.

Tamara Hermosilla1, Daniel Muñoz, Rodrigo Herrera-Molina

  • 1Program of Cell and Molecular Biology, FONDAP, Center for Molecular Studies of the Cell (CEMC), Institute of Biomedical Sciences (ICBM), Facultad de Medicina, Universidad de Chile, Santiago, Chile.

Biochimica Et Biophysica Acta
|March 19, 2008
PubMed
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Neuronal Thy-1 directly interacts with astrocyte alphavbeta3 integrin, triggering signaling pathways essential for neuron-astrocyte communication and focal adhesion formation. This clarifies Thy-1

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Molecular Biology

Background:

  • Thy-1 is an abundant neuronal glycoprotein with an unclear function.
  • Previous studies suggested Thy-1 clusters beta3 integrin in astrocytes, initiating signaling cascades.
  • The alpha subunit partner of beta3 integrin and the mechanism of neuronal Thy-1-induced astrocyte signaling were unknown.

Purpose of the Study:

  • To identify the alpha subunit partner of beta3 integrin in DI TNC1 astrocytes.
  • To investigate the role of the Thy-1-integrin interaction in neuron-astrocyte signaling.
  • To confirm direct interaction between neuronal Thy-1 and astrocyte alphavbeta3 integrin.

Main Methods:

  • Cell association assays using anti-integrin subunit antibodies.
  • Surface Plasmon Resonance (SPR) analysis to detect direct protein interactions.
  • Western blotting to assess tyrosine phosphorylation and RhoA activation.

Main Results:

  • Evidence was obtained for alphavbeta3 integrin heterodimer formation in DI TNC1 astrocytes.
  • Antibodies against alphav or beta3 integrin reduced neuron-astrocyte cell-cell interaction.
  • Anti-Thy-1 antibodies blocked neuron-induced astrocyte stimulation but not binding.
  • Wild-type Thy-1, but not mutated Thy-1, directly interacted with alphavbeta3 integrin via SPR.
  • This interaction was cation-dependent and species-independent.

Conclusions:

  • The alphavbeta3 integrin heterodimer is formed in astrocytes and mediates neuron-astrocyte interactions.
  • Neuronal Thy-1 directly binds to astrocyte alphavbeta3 integrin.
  • This direct interaction is crucial for Thy-1-mediated signaling, leading to focal adhesion formation in astrocytes.