| Chapter title |
Purification and Analysis of Exosomes Released by Mature Cortical Neurons Following Synaptic Activation.
|
|---|---|
| Chapter number | 9 |
| Book title |
Exosomes and Microvesicles
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6728-5_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6726-1, 978-1-4939-6728-5
|
| Authors |
Karine Laulagnier, Charlotte Javalet, Fiona J. Hemming, Rémy Sadoul |
| Editors |
Andrew F Hill |
| 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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 28 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 8 | 29% |
| Student > Doctoral Student | 3 | 11% |
| Researcher | 3 | 11% |
| Student > Bachelor | 2 | 7% |
| Student > Postgraduate | 2 | 7% |
| Other | 2 | 7% |
| Unknown | 8 | 29% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 7 | 25% |
| Agricultural and Biological Sciences | 4 | 14% |
| Neuroscience | 3 | 11% |
| Chemical Engineering | 1 | 4% |
| Physics and Astronomy | 1 | 4% |
| Other | 3 | 11% |
| Unknown | 9 | 32% |