Journal Club: B-Actin mRNA in neurons (Singer)


Basic observation

  • chemical stimulation of neuron, see more B-actin mRNA in axon (2x)
  • also see more B-actin mRNA following gentle protease treatment — propose mRNA was masked
  • do transcription controls and trafficking controls
  • block neural stimulation pathway, also don’t see more B-actin
  • kinetics of response (detected in less than 10 minutes). within 20-40 min has returned to masked state. (half-life tau = 7 min)
  • is this long enough to make bigger spines? Well certainly if you have multiple stimulations
  • is more actin actually made? Use dendra-2-B-actin photo-conversion to detect newly synthesized B=actin.
  • rRNA detection also increases following protease treatment
  • masked B-actin (revealed by protease) associate more the rRNA granuales (revealed by protease).
  • bright ribosomal structures have reduced mobility, become more mobile following cLTP
  • live imaging of B-actin mRNAs observe faster diffusion in stimulated cells.
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