Dec 21, 2012
Produced in the laboratory of Dr. Herbert Geller at the NIH's National Heart, Lung, and Blood Institute, this movie shows a cultured neonatal mouse cerebellar granule neuron transfected with a construct of monomeric RFP-EB3. A round glass coverslip that formed the bottom of a 35 mm Petri dish was first coated with poly-l-lysine (PLL). Eight-day-old mouse cerebellar granule neurons were dissociated, transfected with RFP-EB3, and plated on the coverslip. After 24 hr, the Petri dish was placed in a CO2 incubator on the stage of a Nikon inverted microscope. Images were captured every 5 sec with a 100X objective using the Metamorph Program. The QuickTime movie was made using ImageJ and represents 25 frames. What you will see is a neuron with EB3 speckles moving outward from the soma and distally in each of the processes. Speckles indicate the sites of tubulin polymerization and the rate of movement of speckles represents the polymerization rate.
Growth cones turn at a boundary