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Labeling F-actin Barbed Ends with Rhodamine-actin in Permeabilized Neuronal Growth Cones
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Chapters
- 00:05Title
- 01:49Preparation of Coverslips or “Video” Dishes
- 03:40Preparation of Neuronal Cultures and Media
- 04:53Permeabilization of Neuronal Cultures and Incorporation of Rhodamine-actin onto F-actin Barbed Ends
- 06:13Variations to this Procedure
- 07:35Representative Actin Labeling Results
- 08:47Conclusion
A method to visualize and quantify F-actin barbed ends in neuronal growth cones is described. After culturing neurons on glass coverslips, cells are permeabilized with a saponin-containing solution. Then, a short incubation with the saponin buffer containing rhodamine-actin incorporates fluorescent actin onto free actin barbed ends.
Tags
F-actinBarbed EndsRhodamine-actinPermeabilizedNeuronal Growth ConesAxonsMolecular Guidance CuesGrowth Cone CytoskeletonActin Filament MeshworkActin PolymerizationDynamic RemodelingGuidance CuesActin MonomerPolymerization NucleiChick RetinalDorsal Root Ganglion (DRG)Actin Depolymerizing Factor (ADF/cofilin)
