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The End-Binding Protein EB3 Localizes Points of Microtubule Polymerization in a Neuron

Below is a link to a Quick Time Movie of a cultured neonatal mouse cerebellar granule neuron transfected with a construct of monomeric RFP-EB3.    A round glass coverslip which 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 Quick Time 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. 

Download the Movie
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Dr. Herbert Geller, Head
Developmental Neurobiology, NHLBI, NIH
Building 10, Room 8C-213, 10 Center Drive, MSC-1650
Bethesda, MD 20892-1650
Email: gellerh@nhlbi.nih.gov
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