| Chapter title |
Single-Cell Imaging Techniques for the Real-Time Detection of IP(3) in Live Cells.
|
|---|---|
| Chapter number | 10 |
| Book title |
Calcium Signaling Protocols
|
| Published in |
Methods in molecular biology, January 2013
|
| DOI | 10.1007/978-1-62703-086-1_10 |
| Pubmed ID | |
| Book ISBNs |
978-1-62703-085-4, 978-1-62703-086-1
|
| Authors |
Carl P. Nelson |
| Abstract |
Inositol 1,4,5-trisphosphate (IP(3)) is a ubiquitous second messenger, derived from the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP(2)) by enzymes of the phospholipase C (PLC) family. Binding of IP(3) to its cognate receptor in the endoplasmic reticulum membrane leads to release of Ca(2+) into the cytoplasm, which is involved in the regulation of an array of cellular functions. Traditional techniques for the detection of IP(3) have required the extraction of a large number of cells, with limitations in the time resolution of changes in IP(3) and an inability to obtain detailed information on the dynamics of this second messenger in single cells. Recent progress in this field has led to the development of a number of genetically encoded fluorescent biosensors, which upon recombinant expression are able selectively to detect real-time changes in IP(3) in single live cells. In this chapter, I detail protocols for the expression, visualization (by confocol or fluorescence microscopy), and interpretation of data obtained with such biosensors expressed in mammalian cells. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 10 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 3 | 30% |
| Professor > Associate Professor | 1 | 10% |
| Unknown | 6 | 60% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 2 | 20% |
| Medicine and Dentistry | 1 | 10% |
| Unknown | 7 | 70% |