Phosphoinositide detection at synapses of fixed murine hippocampal neurons
Summary: The minor phospholipid phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is crucial for neurotransmission and has been implicated in Parkinson’s disease. Here, we present a staining protocol for the analysis of activity-dependent changes of PI(4,5)P2 at synapses. We describe steps for stimu...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724001102 |
| _version_ | 1797224114956533760 |
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| author | Svenja Bolz Natalie Kaempf Maria Muehlbauer Delia Löwe Volker Haucke |
| author_facet | Svenja Bolz Natalie Kaempf Maria Muehlbauer Delia Löwe Volker Haucke |
| author_sort | Svenja Bolz |
| collection | DOAJ |
| description | Summary: The minor phospholipid phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is crucial for neurotransmission and has been implicated in Parkinson’s disease. Here, we present a staining protocol for the analysis of activity-dependent changes of PI(4,5)P2 at synapses. We describe steps for stimulating and fixing murine hippocampal neurons, staining with probes for PI(4,5)P2 and a synaptic marker, and analysis by high-resolution microscopy. Our approach gives insights into local PI(4,5)P2 synthesis and turnover at synapses and can be extended to phosphoinositide lipids other than PI(4,5)P2.For complete details on the use and execution of this protocol, please refer to Bolz et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-24T13:47:59Z |
| format | Article |
| id | doaj.art-2fa3e98cb3fa43138274b867c90f4d1e |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-24T13:47:59Z |
| publishDate | 2024-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-2fa3e98cb3fa43138274b867c90f4d1e2024-04-04T05:07:18ZengElsevierSTAR Protocols2666-16672024-06-0152102945Phosphoinositide detection at synapses of fixed murine hippocampal neuronsSvenja Bolz0Natalie Kaempf1Maria Muehlbauer2Delia Löwe3Volker Haucke4Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany; Faculty of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195 Berlin, GermanyLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, GermanyLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, GermanyLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, GermanyLeibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany; Faculty of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195 Berlin, Germany; NeuroCure Cluster of Excellence, Charité Universitätsmedizin Berlin, 10117 Berlin, Germany; Corresponding authorSummary: The minor phospholipid phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is crucial for neurotransmission and has been implicated in Parkinson’s disease. Here, we present a staining protocol for the analysis of activity-dependent changes of PI(4,5)P2 at synapses. We describe steps for stimulating and fixing murine hippocampal neurons, staining with probes for PI(4,5)P2 and a synaptic marker, and analysis by high-resolution microscopy. Our approach gives insights into local PI(4,5)P2 synthesis and turnover at synapses and can be extended to phosphoinositide lipids other than PI(4,5)P2.For complete details on the use and execution of this protocol, please refer to Bolz et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724001102Cell BiologyCell MembraneMicroscopyAntibodyNeuroscienceProtein expression and purification |
| spellingShingle | Svenja Bolz Natalie Kaempf Maria Muehlbauer Delia Löwe Volker Haucke Phosphoinositide detection at synapses of fixed murine hippocampal neurons STAR Protocols Cell Biology Cell Membrane Microscopy Antibody Neuroscience Protein expression and purification |
| title | Phosphoinositide detection at synapses of fixed murine hippocampal neurons |
| title_full | Phosphoinositide detection at synapses of fixed murine hippocampal neurons |
| title_fullStr | Phosphoinositide detection at synapses of fixed murine hippocampal neurons |
| title_full_unstemmed | Phosphoinositide detection at synapses of fixed murine hippocampal neurons |
| title_short | Phosphoinositide detection at synapses of fixed murine hippocampal neurons |
| title_sort | phosphoinositide detection at synapses of fixed murine hippocampal neurons |
| topic | Cell Biology Cell Membrane Microscopy Antibody Neuroscience Protein expression and purification |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724001102 |
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