S'identifier

David Geffen School of Medicine, UCLA

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Study Glial Cell Heterogeneity Influence on Axon Growth Using a New Coculture Method

Han-peng Xu^1,2, Lin Gou^3,4, Hong-Wei Dong³

¹Department of Neurosurgery, Cedars Sinai Medical Center, UCLA, ²Basic Medicine School, Fourth Military Medical University, ³Department of Neurology, David Geffen School of Medicine, UCLA, ⁴Aerospace Medicine School, Fourth Military Medical Univeristy

In this protocol, we described a new method to study the influence of glial cell heterogeneity on axon growth with an in vitro co-culture system. Rat cortical glial cells were cultured to confluence and cocultured with highly purified rat dorsal root ganglia neurons. Different glial cell influence on neurons adhesion and axon growth was compared directly in the same culture. This method provides a new way to directly study the glial cell heterogeneity influence on neuron adhesion and axon growth.

Biology

High-throughput Crystallization of Membrane Proteins Using the Lipidic Bicelle Method

Rachna Ujwal¹, Jeff Abramson²

¹UCLA-DOE Institute for Genomics and Proteomics, University of California Los Angeles , ²Department of Physiology, David Geffen School of Medicine, UCLA

Bicelles are lipid/amphiphile mixtures that maintain membrane proteins (MPs) within a lipid bilayer but have unique phase behavior that facilitates high-throughput screening by crystallization robots. This technique has successfully produced a number of high-resolution structures from both prokaryotic and eukaryotic sources. This video describes protocols for generating the lipidic bicelle mixture, incorporating MPs into the bicelle mixture, setting up crystallizations trials (manually as well as robotically) and harvesting crystals from the medium.