| RRC ID |
52003
|
| 著者 |
Niwa F, Sakuragi S, Kobayashi A, Takagi S, Oda Y, Bannai H, Mikoshiba K.
|
| タイトル |
Dissection of local Ca(2+) signals inside cytosol by ER-targeted Ca(2+) indicator.
|
| ジャーナル |
Biochem Biophys Res Commun
|
| Abstract |
Calcium (Ca(2+)) is a versatile intracellular second messenger that operates in various signaling pathways leading to multiple biological outputs. The diversity of spatiotemporal patterns of Ca(2+) signals, generated by the coordination of Ca(2+) influx from the extracellular space and Ca(2+) release from the intracellular Ca(2+) store the endoplasmic reticulum (ER), is considered to underlie the diversity of biological outputs caused by a single signaling molecule. However, such Ca(2+) signaling diversity has not been well described because of technical limitations. Here, we describe a new method to report Ca(2+) signals at subcellular resolution. We report that OER-GCaMP6f, a genetically encoded Ca(2+) indicator (GECI) targeted to the outer ER membrane, can monitor Ca(2+) release from the ER at higher spatiotemporal resolution than conventional GCaMP6f. OER-GCaMP6f was used for in vivo Ca(2+) imaging of C. elegans. We also found that the spontaneous Ca(2+) elevation in cultured astrocytes reported by OER-GCaMP6f showed a distinct spatiotemporal pattern from that monitored by plasma membrane-targeted GCaMP6f (Lck-GCaMP6f); less frequent Ca(2+) signal was detected by OER-GCaMP6f, in spite of the fact that Ca(2+) release from the ER plays important roles in astrocytes. These findings suggest that targeting of GECIs to the ER outer membrane enables sensitive detection of Ca(2+) release from the ER at subcellular resolution, avoiding the diffusion of GECI and Ca(2+). Our results indicate that Ca(2+) imaging with OER-GCaMP6f in combination with Lck-GCaMP6f can contribute to describing the diversity of Ca(2+) signals, by enabling dissection of Ca(2+) signals at subcellular resolution.
|
| 巻・号 |
479(1)
|
| ページ |
67-73
|
| 公開日 |
2016-10-7
|
| DOI |
10.1016/j.bbrc.2016.09.034
|
| PII |
S0006-291X(16)31485-1
|
| PMID |
27616195
|
| MeSH |
Animals
Animals, Genetically Modified
Astrocytes / cytology
Astrocytes / metabolism
COS Cells
Caenorhabditis elegans / genetics
Caenorhabditis elegans / metabolism
Calcium / metabolism*
Calcium Signaling*
Cell Membrane / metabolism
Cells, Cultured
Chlorocebus aethiops
Cytosol / metabolism*
Endoplasmic Reticulum / metabolism*
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism*
HeLa Cells
Humans
Microscopy, Confocal
Rats, Wistar
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Time-Lapse Imaging / methods
|
| IF |
2.985
|
| 引用数 |
8
|
| リソース情報 |
| ヒト・動物細胞 |
COS-7(RCB0539)
HeLa(RCB0007) |
