論文 - 詳細
| RRC ID | 54924 |
|---|---|
| 著者 | Bannai H, Hirose M, Niwa F, Mikoshiba K. |
| タイトル | Dissection of Local Ca2+ Signals in Cultured Cells by Membrane-targeted Ca2+ Indicators. |
| ジャーナル | J Vis Exp |
| Abstract |
Calcium ion (Ca2+) is a universal intracellular messenger molecule that drives multiple signaling pathways, leading to diverse biological outputs. The coordination of two Ca2+ signal sources-"Ca2+ influx" from outside the cell and "Ca2+ release" from the intracellular Ca2+ store endoplasmic reticulum (ER)-is considered to underlie the diverse spatio-temporal patterns of Ca2+ signals that cause multiple biological functions in cells. The purpose of this protocol is to describe a new Ca2+ imaging method that enables monitoring of the very moment of "Ca2+ influx" and "Ca2+ release". OER-GCaMP6f is a genetically encoded Ca2+ indicator (GECI) comprising GCaMP6f, which is targeted to the ER outer membrane. OER-GCaMP6f can monitor Ca2+ release at a higher temporal resolution than conventional GCaMP6f. Combined with plasma membrane-targeted GECIs, the spatio-temporal Ca2+ signal pattern can be described at a subcellular resolution. The subcellular-targeted Ca2+ indicators described here are, in principle, available for all cell types, even for the in vivo imaging of Caenorhabditis elegans neurons. In this protocol, we introduce Ca2+ imaging in cells from cell lines, neurons, and glial cells in dissociated primary cultures, and describe the preparation of frozen stock of rat cortical neurons. |
| 巻・号 | (145) |
| 公開日 | 2019-3-22 |
| DOI | 10.3791/59246 |
| PMID | 30958464 |
| MeSH | Animals Caenorhabditis elegans / cytology Calcium / metabolism* Calcium Signaling* Cell Membrane / metabolism* Cells, Cultured Endoplasmic Reticulum / metabolism Neurons / cytology Rats |
| IF | 1.108 |
| 引用数 | 3 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 21 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HeLa(RCB0007) |