- 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.