PICK1 interacts with α7 neuronal nicotinic acetylcholine receptors and controls their clustering

Baer, Kristin; Bürli, Thomas; Huh, Kyung-Hye; Wiesner, Andreas; Erb-Vögtli, Susanne; Göckeritz-Dujmovic, Dubravka; Moransard, Martijn; Nishimune, Atsushi; Rees, Mark I.; Henley, Jeremy M.; Fritschy, Jean‐Marc; Fuhrer, Christian

doi:10.1016/j.mcn.2007.03.009

Cited by 34 publications

(53 citation statements)

References 83 publications

(169 reference statements)

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“…These include promoting clustering of the binding partners in cellular subcompartments (8,10,(15)(16)(17), regulation of their trafficking (10, 18 -20), and acting as a scaffold that brings PKC␣ in close proximity to other binding partners to govern their phosphorylation (9,(21)(22)(23).…”

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confidence: 99%

Protein Interacting with C Kinase 1 (PICK1) Reduces Reinsertion Rates of Interaction Partners Sorted to Rab11-dependent Slow Recycling Pathway

Madsen

Thorsen

Rahbek-Clemmensen

et al. 2012

Journal of Biological Chemistry

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Background:The role of PICK1 in regulating trafficking of its PDZ domain binding partners (e.g. AMPA receptors) remains unclear. Results: PICK1 clusters and reduces recycling only of PDZ binding partners sorted to Rab11-dependent recycling. Conclusion: Contrary to other PDZ domain proteins, which regulate postendocytic sorting, PICK1 determines the trafficking rate through an endocytic compartment. Significance: This function might explain the role of PICK1 in synaptic plasticity.

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confidence: 99%

Protein Interacting with C Kinase 1 (PICK1) Reduces Reinsertion Rates of Interaction Partners Sorted to Rab11-dependent Slow Recycling Pathway

Madsen

Thorsen

Rahbek-Clemmensen

et al. 2012

Journal of Biological Chemistry

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“…One candidate for the link would be the PDZ-containing scaffold protein PICK1. PICK1 has recently been shown to bind to α7-nAChRs (Baer et al, 2007), and EphB2Rs are thought to bind PICK1 as well (Torres et al, 1998). PICK1 is usually not found concentrated at synapses, however, and in the case of AMPA receptors is thought to participate in trafficking both to and from the surface, rather than synaptic stabilization (Lu and Ziff, 2005;Cao et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

EphB Receptors Co-Distribute with a Nicotinic Receptor Subtype and Regulate Nicotinic Downstream Signaling in Neurons

Liu¹,

Conroy²,

Stawicki³

et al. 2008

Molecular and Cellular Neuroscience

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Activation of nicotinic acetylcholine receptors (nAChRs) on neurons engages calcium-dependent signaling pathways regulating numerous events. Receptors containing α7 subunits (α7-nAChRs) are prominent in this because of their abundance and high relative calcium permeability. We show here that EphB2 receptors are co-localized with postsynaptic α7-nAChRs on chick ciliary ganglion neurons and that treatment of the cells with an ephrinB1 construct to activate the EphB receptors exerts physical restraints on both classes of receptors, diminishing their dispersal after spine retraction or lipid raft disruption. Moreover, the ephrinB1/EphB receptor complex specifically enhances the ability of α7-nAChRs to activate the transcription factor CREB, acting through a pathway including a receptor tyrosine kinase, a Src family member, PI3 kinase, and protein kinase A most distally. The enhancement does not appear to result from a change in the α7-nAChR current amplitude, suggesting a downstream target. The results demonstrate a role for ephrin/EphB action in nicotinic signaling.

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“…The proteins chosen were a7 nAChR, a nicotinic acetylcholine receptor found in many neurons throughout the CNS and particularly enriched in GABAergic interneurons in the hippocampus, and PICK1, a protein involved in the trafficking of glutamate receptors and enriched at glutamatergic synapses 10,19,20 . Our method achieved expression of these two genes in pyramidal hippocampal neurons and even in a small subpool of cells, the hippocampal GABAergic interneurons.…”

Section: Application 2: Double-magnetofection Of Gabaergic Interneuromentioning

confidence: 99%

“…The slow development of GABAergic synapses proceeded normally under these circumstances, showing that gephyrin-GFP expression did not interfere with neural development and synapse formation. In the second application, we concentrated on two genes, a7 nicotinic acetylcholine receptor and the scaffold protein PICK1 10 . We achieved simultaneous expression of these two genes in GABAergic interneurons, which form a small subset of cells.…”

Section: Introductionmentioning

confidence: 99%

“…This protocol shows low toxicity, uses a combination of a transfection reagent, CombiMag, and magnetic force and shows a sufficiently high efficiency in neurons (ca. 5%) to allow single-cell analysis.We present a core protocol and illustrate the critical steps, modifications and applications carried out by our three laboratories (Medina, Fuhrer and Fritschy) [8][9][10] . This protocol allows single and double transfections of DNA vectors at different time points after plating.…”

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Efficient transfection of DNA or shRNA vectors into neurons using magnetofection

et al. 2007

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Efficient and long-lasting transfection of primary neurons is an essential tool for addressing many questions in current neuroscience using functional gene analysis. Neurons are sensitive to cytotoxicity and difficult to transfect with most methods. We provide a protocol for transfection of cDNA and RNA interference (short hairpin RNA (shRNA)) vectors, using magnetofection, into rat hippocampal neurons (embryonic day 18/19) cultured for several hours to 21 d in vitro. This protocol even allows double-transfection of DNA into a small subpopulation of hippocampal neurons (GABAergic interneurons), as well as achieving long-lasting expression of DNA and shRNA constructs without interfering with neuronal differentiation. This protocol, which uses inexpensive equipment and reagents, takes 1 h; utilizes mixed hippocampal cultures, a transfection reagent, CombiMag, and a magnetic plate; shows low toxicity and is suited for single-cell analysis. Modifications done by our three laboratories are detailed. INTRODUCTIONEvery gene transfer method aims to achieve high transfection efficiency, low toxicity and long-lasting expression. These goals are especially difficult to achieve for postmitotic primary neurons due to their high sensitivity to any microenvironmental change 1-5 . Many currently available transfection methods do not yield sufficiently good gene expression results to allow functional gene analysis in differentiated adherent neurons in vitro. A recent report provides a useful overview of seven different gene delivery methods, including two often-used transfection techniques (calcium-phosphate and lipofection), but excluding magnetofection 6 . Magnetofection is a technique that can be used to reliably and efficiently introduce DNA into a variety of cell types 7 . Therefore, we have invested in designing a reliable, magnetofection-based protocol to express DNA or shRNA constructs in rat hippocampal neurons (embryonic day (E)18/19) cultured for several hours to 21 d in vitro (DIV). After transfection, neurons expressing exogenous proteins can develop normally in culture for 5-10 d. We have determined the optimum parameters for transfection efficiency such as age of cells at transfection time point, expression levels during the time after transfection and parameters for doubletransfection. This protocol shows low toxicity, uses a combination of a transfection reagent, CombiMag, and magnetic force and shows a sufficiently high efficiency in neurons (ca. 5%) to allow single-cell analysis.We present a core protocol and illustrate the critical steps, modifications and applications carried out by our three laboratories (Medina, Fuhrer and Fritschy) [8][9][10] . This protocol allows single and double transfections of DNA vectors at different time points after plating. First, we present a detailed characterization of magnetofection parameters such as age of neurons, time course of protein expression after magnetofection, and transfection efficiency in pyramidal neurons and GABAergic interneurons. Three specific

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PICK1 interacts with α7 neuronal nicotinic acetylcholine receptors and controls their clustering

Cited by 34 publications

References 83 publications

Protein Interacting with C Kinase 1 (PICK1) Reduces Reinsertion Rates of Interaction Partners Sorted to Rab11-dependent Slow Recycling Pathway

Protein Interacting with C Kinase 1 (PICK1) Reduces Reinsertion Rates of Interaction Partners Sorted to Rab11-dependent Slow Recycling Pathway

EphB Receptors Co-Distribute with a Nicotinic Receptor Subtype and Regulate Nicotinic Downstream Signaling in Neurons

Efficient transfection of DNA or shRNA vectors into neurons using magnetofection

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