| Chapter title |
Imaging the Polarized Sorting of Proteins from the Golgi Complex in Live Neurons
|
|---|---|
| Chapter number | 2 |
| Book title |
The Golgi Complex
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-6463-5_2 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6461-1, 978-1-4939-6463-5
|
| Authors |
Ginny G. Farías, Dylan J. Britt, Juan S. Bonifacino |
| Editors |
William J. Brown |
| Abstract |
The study of polarized protein trafficking in live neurons is critical for understanding neuronal structure and function. Given the complex anatomy of neurons and the numerous trafficking pathways that are active in them, however, visualization of specific vesicle populations leaving the Golgi complex presents unique challenges. Indeed, several approaches used in non-polarized cells, and even in polarized epithelial cells, have been less successful in neurons. Here, we describe an adaptation of the recently developed Retention Using Selective Hooks (RUSH) system (Boncompain et al., Nat Methods 9:493-498, 2012), previously used in non-polarized cells, to analyze the polarized sorting of proteins from the Golgi complex to dendrites and axons in live neurons. The RUSH system involves the retention of a fluorescently tagged cargo protein fused to the streptavidin-binding peptide (SBP) in the endoplasmic reticulum (ER) through the expression of an ER-hook protein fused to streptavidin. Upon D-biotin addition, the cargo protein is released and its traffic to dendrites and axons can be analyzed in live neurons. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 20% |
| United States | 1 | 20% |
| Brazil | 1 | 20% |
| Spain | 1 | 20% |
| France | 1 | 20% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 3 | 60% |
| Science communicators (journalists, bloggers, editors) | 1 | 20% |
| Members of the public | 1 | 20% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 46 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 13 | 28% |
| Researcher | 11 | 24% |
| Professor | 2 | 4% |
| Student > Bachelor | 2 | 4% |
| Student > Postgraduate | 2 | 4% |
| Other | 6 | 13% |
| Unknown | 10 | 22% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 13 | 28% |
| Agricultural and Biological Sciences | 12 | 26% |
| Neuroscience | 11 | 24% |
| Economics, Econometrics and Finance | 1 | 2% |
| Unspecified | 1 | 2% |
| Other | 0 | 0% |
| Unknown | 8 | 17% |