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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Holz, R. W. Micheva, K. D. Smith, S. J. |
| Description | Author Affiliation: Micheva KD ( Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305, USA. kmicheva@stanford.edu) |
| Abstract | Phosphatidylinositol 4,5-biphosphate (PIP2) has been implicated in a variety of cellular processes, including synaptic vesicle recycling. However, little is known about the spatial distribution of this phospholipid in neurons and its dynamics. In this study, we have focused on these questions by transiently expressing the phospholipase C (PLC)-δ1 pleckstrin homology (PH) domain fused to green fluorescent protein (GFP) in cultured hippocampal neurons. This PH domain binds specifically and with high affinity to PIP2. Live confocal imaging revealed that in resting cells, PH-GFP is localized predominantly on the plasma membrane. Interestingly, no association of PH-GFP with synaptic vesicles in quiescent neurons was observed, indicating the absence of detectable PIP2 on mature synaptic vesicles. Electrical stimulation of hippocampal neurons resulted in a decrease of the PH-GFP signal at the plasma membrane, most probably due to a PLC-mediated hydrolysis of PIP2. This was accompanied in the majority of presynaptic terminals by a marked increase in the cytoplasmic PH-GFP signal, localized most probably on freshly endocytosed membranes. Further investigation revealed that the increase in PH-GFP signal was dependent on the activation of N-methyl-D-aspartate receptors and the consequent production of nitric oxide (NO). Thus, PIP2 in the presynaptic terminal appears to be regulated by postsynaptic activity via a retrograde action of NO. |
| ISSN | 00219525 |
| e-ISSN | 15408140 |
| Journal | The Journal of Cell Biology |
| Issue Number | 2 |
| Volume Number | 154 |
| Language | English |
| Publisher | Rockefeller University Press (United States) |
| Publisher Date | 2001-07-23 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Neurons Enzymology Phosphatidylinositol 4,5-Diphosphate Metabolism Presynaptic Terminals Animals Blood Proteins Genetics Cell Membrane Cell Survival Drug Effects Cells, Cultured Electric Stimulation Endocytosis Enzyme Inhibitors Pharmacology Exocytosis Green Fluorescent Proteins Hippocampus Isoenzymes Antagonists & Inhibitors Luminescent Proteins Microscopy, Fluorescence Cytology Nitric Oxide Biosynthesis Phospholipase C Delta Phosphoproteins Protein Structure, Tertiary Physiology Receptors, N-Methyl-D-Aspartate Recombinant Fusion Proteins Synaptic Vesicles Transfection Type C Phospholipases Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Cell Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine |
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