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University of Lyon 1

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Use of pHluorin to Assess the Dynamics of Axon Guidance Receptors in Cell Culture and in the Chick Embryo

Céline Delloye-Bourgeois^*1, Arnaud Jacquier^*1, Julien Falk¹, Valérie Castellani¹

¹University Claude Bernard Lyon1, CGphiMC UMR CNRS 5534, University of Lyon

We describe here the use of a pH-sensitive green fluorescent protein variant, pHluorin, to study the spatio-temporal dynamics of axon guidance receptors trafficking at the cell surface. The pHluorin-tagged receptor is expressed both in cell culture and in vivo, using electroporation of the chick embryo.

Neuroscience

Culture of Isolated Floor Plate Tissue and Production of Conditioned Medium to Assess Functional Properties of Floor Plate-released Signals

Camille Charoy¹, Elise Arbeille¹, Karine Thoinet¹, Valérie Castellani¹

¹CGphiMC, UMR CNRS 5534, University of Lyon 1

The floor plate is a crucial structure of the developing spinal cord, providing diffusible signals to progenitors and differentiating neurons. We describe a method to produce floor-plate conditioned medium, to apply it to fresh spinal cord tissue, and to assess the consequences on proteins of interest by biochemistry.