Article ; Online: TRPM7 facilitates cholinergic vesicle fusion with the plasma membrane.
Proceedings of the National Academy of Sciences of the United States of America
2008 Volume 105, Issue 24, Page(s) 8304–8308
Abstract: ... plasma membrane fusion reporter. In PC12 cells, as in sympathetic neurons, TRPM7 is located in ACh-secreting SSLVs ... without affecting large dense core vesicle secretion. We conclude that the conductance of TRPM7 across the vesicle ... membrane is important in SSLV fusion. ...
| Abstract | TRPM7, of the transient receptor potential (TRP) family, is both an ion channel and a kinase. Previously, we showed that TRPM7 is located in the membranes of acetylcholine (ACh)-secreting synaptic vesicles of sympathetic neurons, forms a molecular complex with proteins of the vesicular fusion machinery, and is critical for stimulated neurotransmitter release. Here, we targeted pHluorin to small synaptic-like vesicles (SSLV) in PC12 cells and demonstrate that it can serve as a single-vesicle plasma membrane fusion reporter. In PC12 cells, as in sympathetic neurons, TRPM7 is located in ACh-secreting SSLVs. TRPM7 knockdown by siRNA, or abolishing channel activity by expression of a dominant negative TRPM7 pore mutant, decreased the frequency of spontaneous and voltage-stimulated SSLV fusion events without affecting large dense core vesicle secretion. We conclude that the conductance of TRPM7 across the vesicle membrane is important in SSLV fusion. |
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| MeSH term(s) | Acetylcholine/metabolism ; Animals ; Biological Transport ; Cell Membrane/enzymology ; Cell Membrane/physiology ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Membrane Fusion ; PC12 Cells ; Phosphotransferases/genetics ; Phosphotransferases/metabolism ; RNA, Small Interfering/genetics ; Rats ; Synaptic Vesicles/enzymology ; Synaptic Vesicles/physiology ; TRPM Cation Channels/genetics ; TRPM Cation Channels/metabolism ; Vesicular Acetylcholine Transport Proteins/genetics ; Vesicular Acetylcholine Transport Proteins/metabolism |
| Chemical Substances | PHluorin ; RNA, Small Interfering ; TRPM Cation Channels ; Vesicular Acetylcholine Transport Proteins ; Green Fluorescent Proteins (147336-22-9) ; Phosphotransferases (EC 2.7.-) ; Trpm7 protein, rat (EC 2.7.11.1) ; Acetylcholine (N9YNS0M02X) |
| Language | English |
| Publishing date | 2008-06-06 |
| Publishing country | United States |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 209104-5 |
| ISSN | 1091-6490 ; 0027-8424 |
| ISSN (online) | 1091-6490 |
| ISSN | 0027-8424 |
| DOI | 10.1073/pnas.0800881105 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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