NDLI: Neural Explant Cultures from Xenopus laevis.

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Neural Explant Cultures from Xenopus laevis.

Year: 2012 Issue: 67

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Neural Explant Cultures from Xenopus laevis.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lowery, Laura Anne Faris, Anna E. R. Stout, Alina Van Vactor, David
Description	Country affiliation: United States Author Affiliation: Lowery LA ( Department of Cell Biology, Harvard Medical School, Boston, MA, USA. Laura_Lowery@hms.harvard.edu)
Abstract	The complex process of axon guidance is largely driven by the growth cone, which is the dynamic motile structure at the tip of the growing axon. During axon outgrowth, the growth cone must integrate multiple sources of guidance cue information to modulate its cytoskeleton in order to propel the growth cone forward and accurately navigate to find its specific targets(1). How this integration occurs at the cytoskeletal level is still emerging, and examination of cytoskeletal protein and effector dynamics within the growth cone can allow the elucidation of these mechanisms. Xenopus laevis growth cones are large enough (10-30 microns in diameter) to perform high-resolution live imaging of cytoskeletal dynamics (e.g.(2-4) ) and are easy to isolate and manipulate in a lab setting compared to other vertebrates. The frog is a classic model system for developmental neurobiology studies, and important early insights into growth cone microtubule dynamics were initially found using this system(5-7) . In this method(8), eggs are collected and fertilized in vitro, injected with RNA encoding fluorescently tagged cytoskeletal fusion proteins or other constructs to manipulate gene expression, and then allowed to develop to the neural tube stage. Neural tubes are isolated by dissection and then are cultured, and growth cones on outgrowing neurites are imaged. In this article, we describe how to perform this method, the goal of which is to culture Xenopus laevis growth cones for subsequent high-resolution image analysis. While we provide the example of +TIP fusion protein EB1-GFP, this method can be applied to any number of proteins to elucidate their behaviors within the growth cone.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/4232
Journal	Journal of Visualized Experiments
Issue Number	68
Language	English
Publisher	MyJove Corp.
Publisher Date	2012-10-15
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Growth Cones Ultrastructure Neural Tube Growth & Development Neurites Organ Culture Techniques Animals Xenopus Laevis Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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