Purification and Analysis of Exosomes Released by Mature Cortical Neurons Following Synaptic Activation.
Methods Mol Biol
; 1545: 129-138, 2017.
Article
en En
| MEDLINE
| ID: mdl-27943211
ABSTRACT
Exosomes are vesicles released by most cells into their environment upon fusion of multivesicular endosomes with the plasma membrane. Exosomes are vesicles of 60-100 nm in diameter, floating in sucrose at a density of ~1.15 g/mL and carrying a number of marker proteins such as Alix, Tsg101, and Flotillin-1. We use dissociated cortical neurons cultured for around two weeks as exosome-releasing cells. In these conditions, neurons make mature synapses and form networks that can be activated by physiological stimuli. Here, we describe methods to culture differentiated cortical neurons, induce exosome release by increasing glutamatergic synapse activity, and purify exosomes by differential centrifugations followed by density separation using sucrose gradients. These protocols allow purification of neuronal exosomes released within minutes of activation of glutamatergic synapses.
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Bases de datos:
MEDLINE
Asunto principal:
Vesículas Sinápticas
/
Fraccionamiento Celular
/
Corteza Cerebral
/
Exosomas
/
Neuronas
Límite:
Animals
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2017
Tipo del documento:
Article
País de afiliación:
Francia