scholarly journals Selective dendritic localization of mRNA in Drosophila mushroom body output neurons

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Jessica Mitchell ◽  
Carlas S Smith ◽  
Josh Titlow ◽  
Nils Otto ◽  
Pieter van Velde ◽  
...  
Keyword(s):  
Single Molecule ◽  
Nicotinic Acetylcholine Receptors ◽  
Neuronal Plasticity ◽  
Mushroom Body ◽  
Acetylcholine Receptors ◽  
Behavioural Change ◽  
Before And After ◽  
Specific Mrnas ◽  
Output Neurons ◽  
Subcellular Resolution

Memory-relevant neuronal plasticity is believed to require local translation of new proteins at synapses. Understanding this process requires the visualization of the relevant mRNAs within these neuronal compartments. Here we used single-molecule fluorescence in situ hybridization (smFISH) to localize mRNAs at subcellular resolution in the adult Drosophila brain. mRNAs for subunits of nicotinic acetylcholine receptors and kinases could be detected within the dendrites of co-labelled Mushroom Body Output Neurons (MBONs) and their relative abundance showed cell-specificity. Moreover, aversive olfactory learning produced a transient increase in the level of CaMKII mRNA within the dendritic compartments of the 52a MBONs. Localization of specific mRNAs in MBONs before and after learning represents a critical step towards deciphering the role of dendritic translation in the neuronal plasticity underlying behavioural change in Drosophila.

scholarly journals Dendritic localization of mRNA in Drosophila Mushroom Body Output Neurons

2020 ◽  
Author(s):  
Jessica Mitchell ◽  
Carlas S. Smith ◽  
Josh Titlow ◽  
Nils Otto ◽  
Pieter van Velde ◽  
...  
Keyword(s):  
Single Molecule ◽  
Nicotinic Acetylcholine Receptors ◽  
Neuronal Plasticity ◽  
Mushroom Body ◽  
Acetylcholine Receptors ◽  
Behavioural Change ◽  
Before And After ◽  
Specific Mrnas ◽  
Output Neurons ◽  
Subcellular Resolution

AbstractMemory-relevant neuronal plasticity is believed to require local translation of new proteins at synapses. Understanding this process requires the visualization of the relevant mRNAs within these neuronal compartments. Here we used single-molecule fluorescence in situ hybridization (smFISH) to localize mRNAs at subcellular resolution in the adult Drosophila brain. mRNAs for subunits of nicotinic acetylcholine receptors and kinases could be detected within the dendrites of co-labelled Mushroom Body Output Neurons (MBONs) and their relative abundance showed cell-specificity. Moreover, aversive olfactory learning produced a transient increase in the level of CaMKII mRNA within the dendritic compartments of the γ5β′2a MBONs. Localization of specific mRNAs in MBONs before and after learning represents a critical step towards deciphering the role of dendritic translation in the neuronal plasticity underlying behavioural change in Drosophila.

scholarly journals Single Molecule Motion Maps of Open and Desensitization States of Nicotinic Acetylcholine Receptors

2014 ◽  
Vol 106 (2) ◽  
pp. 629a
Author(s):  
Hiroshi Sekiguchi ◽  
Maki Tokue ◽  
Yuri Nishino ◽  
Kouhei Ichiyanagi ◽  
Naoto Yagi ◽  
...  
Keyword(s):  
Single Molecule ◽  
Nicotinic Acetylcholine Receptors ◽  
Acetylcholine Receptors ◽  
Nicotinic Acetylcholine

scholarly journals Transneuronal Dpr12/DIP-δ interactions facilitate compartmentalized dopaminergic innervation of Drosophila mushroom body axons

2019 ◽  
Author(s):  
Bavat Bornstein ◽  
Idan Alyagor ◽  
Victoria Berkun ◽  
Hagar Meltzer ◽  
Fabienne Reh ◽  
...  
Keyword(s):  
Mushroom Body ◽  
Immunoglobulin Superfamily ◽  
Cellular Mechanisms ◽  
Interacting Protein ◽  
Zone Formation ◽  
Unique System ◽  
Output Neurons ◽  
Subcellular Resolution ◽  
Circuit Formation ◽  
Immunoglobulin Superfamily Protein

SummaryThe mechanisms controlling wiring of neuronal networks are largely unknown. The stereotypic architecture of the Drosophila mushroom-body (MB) offers a unique system to study circuit assembly. The adult medial MB γ-lobe is comprised of a long bundle of axons that wires with specific modulatory and output neurons in a tiled manner defining five distinct zones. We found that the immunoglobulin superfamily protein Dpr12 is cell-autonomously required in γ-neurons for their developmental regrowth into the distal γ4/5 zones, where both Dpr12 and its interacting protein, DIP-δ, are enriched. DIP-δ functions in a subset of dopaminergic neurons that wire with γ-neurons within the γ4/5 zone. During metamorphosis, these dopaminergic projections arrive to the γ4/5 zone prior to γ-axons, suggesting that γ-axons extend through a prepatterned region. Thus, Dpr12/DIP-δ transneuronal interaction is required for γ4/5 zone formation. Our study sheds light onto molecular and cellular mechanisms underlying circuit formation within subcellular resolution.

scholarly journals Inhibitory muscarinic acetylcholine receptors enhance aversive olfactory conditioning in adult Drosophila

2018 ◽  
Author(s):  
Noa Bielopolski ◽  
Hoger Amin ◽  
Anthi A. Apostolopoulou ◽  
Eyal Rozenfeld ◽  
Hadas Lerner ◽  
...  
Keyword(s):  
Mushroom Body ◽  
Acetylcholine Receptors ◽  
Physiological Function ◽  
Muscarinic Acetylcholine Receptors ◽  
Kenyon Cell ◽  
Olfactory Conditioning ◽  
Muscarinic Acetylcholine ◽  
Kenyon Cells ◽  
Output Neurons ◽  
Adult Drosophila

AbstractOlfactory associative learning inDrosophilais mediated by synaptic plasticity between the Kenyon cells of the mushroom body and their output neurons. Both Kenyon cells and their inputs are cholinergic, yet little is known about the physiological function of muscarinic acetylcholine receptors in learning in adult flies. Here we show that aversive olfactory learning in adult flies requires type A muscarinic acetylcholine receptors (mAChR-A) specifically in the gamma subtype of Kenyon cells. Surprisingly, mAChR-A inhibits odor responses in both Kenyon cell dendrites and axons. Moreover, mAChR-A knockdown impairs the learning-associated depression of odor responses in a mushroom body output neuron. Our results suggest that mAChR-A is required at Kenyon cell presynaptic terminals to depress the synapses between Kenyon cells and their output neurons, and may suggest a role for the recently discovered axo-axonal synapses between Kenyon cells.

scholarly journals Single-Molecule Imaging of AMPA and Nicotinic Acetylcholine Receptors in Mammalian Cells

2009 ◽  
Vol 96 (3) ◽  
pp. 29a
Author(s):  
Paul D. Simonson ◽  
John Alexander ◽  
Okunola Jeyifous ◽  
William N. Green ◽  
Paul R. Selvin
Keyword(s):  
Single Molecule ◽  
Nicotinic Acetylcholine Receptors ◽  
Mammalian Cells ◽  
Acetylcholine Receptors ◽  
Single Molecule Imaging ◽  
Nicotinic Acetylcholine

scholarly journals Inhibitory muscarinic acetylcholine receptors enhance aversive olfactory learning in adult Drosophila

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Noa Bielopolski ◽  
Hoger Amin ◽  
Anthi A Apostolopoulou ◽  
Eyal Rozenfeld ◽  
Hadas Lerner ◽  
...  
Keyword(s):  
Synaptic Plasticity ◽  
Mushroom Body ◽  
Acetylcholine Receptors ◽  
Olfactory Learning ◽  
Muscarinic Acetylcholine Receptors ◽  
Projection Neurons ◽  
Kenyon Cell ◽  
Muscarinic Acetylcholine ◽  
Kenyon Cells ◽  
Output Neurons

Olfactory associative learning in Drosophila is mediated by synaptic plasticity between the Kenyon cells of the mushroom body and their output neurons. Both Kenyon cells and their inputs from projection neurons are cholinergic, yet little is known about the physiological function of muscarinic acetylcholine receptors in learning in adult flies. Here, we show that aversive olfactory learning in adult flies requires type A muscarinic acetylcholine receptors (mAChR-A), particularly in the gamma subtype of Kenyon cells. mAChR-A inhibits odor responses and is localized in Kenyon cell dendrites. Moreover, mAChR-A knockdown impairs the learning-associated depression of odor responses in a mushroom body output neuron. Our results suggest that mAChR-A function in Kenyon cell dendrites is required for synaptic plasticity between Kenyon cells and their output neurons.

scholarly journals The Effects of Structural Alterations in the Polyamine and Amino Acid Moieties of Philanthotoxins on Nicotinic Acetylcholine Receptor Inhibition in the Locust, Schistocerca gregaria

Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 7007
Author(s):  
Victoria L. Luck ◽  
David P. Richards ◽  
Ashif Y. Shaikh ◽  
Henrik Franzyk ◽  
Ian R. Mellor
Keyword(s):  
Amino Acid ◽  
Nicotinic Acetylcholine Receptors ◽  
Mushroom Body ◽  
Acetylcholine Receptors ◽  
Schistocerca Gregaria ◽  
Dependent Manner ◽  
Nicotinic Acetylcholine ◽  
Whole Cell Patch Clamp ◽  
Receptor Inhibition ◽  
Methylene Groups

Alterations in the polyamine and amino acid (tyrosine) moieties of philanthotoxin-343 (PhTX-343) were investigated for their effects on the antagonism of nicotinic acetylcholine receptors (nAChRs) isolated from the locust (Schistocerca gregaria) mushroom body. Through whole-cell patch-clamp recordings, the philanthotoxin analogues in this study were shown to cause inhibition of the inward current when co-applied with acetylcholine (ACh). PhTX-343 (IC50 = 0.80 μM at −75 mV) antagonised locust nAChRs in a use-dependent manner, suggesting that it acts as an open-channel blocker. The analogue in which both the secondary amine functionalities were replaced with methylene groups (i.e., PhTX-12) was ~6-fold more potent (IC50 (half-maximal inhibitory concentration) = 0.13 μM at −75 mV) than PhTX-343. The analogue containing cyclohexylalanine as a substitute for the tyrosine moiety of PhTX-343 (i.e., Cha-PhTX-343) was also more potent (IC50 = 0.44 μM at −75 mV). A combination of both alterations to PhTX-343 generated the most potent analogue, i.e., Cha-PhTX-12 (IC50 = 1.71 nM at −75 mV). Modulation by PhTX-343 and Cha-PhTX-343 fell into two distinct groups, indicating the presence of two pharmacologically distinct nAChR groups in the locust mushroom body. In the first group, all concentrations of PhTX-343 and Cha-PhTX-343 inhibited responses to ACh. In the second group, application of PhTX-343 or Cha-PhTX-343 at concentrations ≤100 nM caused potentiation, while concentrations ≥1 μM inhibited responses to ACh. Cha-PhTX-12 may have potential to be developed into insecticidal compounds with a novel mode of action.

scholarly journals Antibody-induced crosslinking and cholesterol-sensitive, anomalous diffusion of nicotinic acetylcholine receptors

2019 ◽  
Author(s):  
Alejo Mosqueira ◽  
Pablo A. Camino ◽  
Francisco J. Barrantes
Keyword(s):  
Single Molecule ◽  
Anomalous Diffusion ◽  
Nicotinic Acetylcholine Receptors ◽  
Acetylcholine Receptors ◽  
Dynamic Equilibrium ◽  
Switching Behavior ◽  
Live Cells ◽  
Cholesterol Depletion ◽  
Antigenic Modulation ◽  
Nicotinic Acetylcholine

AbstractSynaptic strength depends on the number of cell-surface neurotransmitter receptors in dynamic equilibrium with intracellular pools. Dysregulation of this homeostatic balance occurs e.g. in myasthenia gravis, an autoimmune disease characterized by a decrease in the number of postsynaptic nicotinic acetylcholine receptors (nAChRs). Monoclonal antibody mAb35 mimics this effect. Here we use STORM nanoscopy to characterize the individual and ensemble dynamics of mAb35-crosslinked receptors in the clonal cell line CHO-K1/A5, which robustly expresses adult muscle-type nAChRs. Antibody labeling of live cells results in 80% receptor immobilization. The remaining mobile fraction exhibits a heterogeneous combination of Brownian and anomalous diffusion. Single-molecule trajectories exhibit a two-state switching behavior between free Brownian walks and anticorrelated walks within confinement areas. The latter act as permeable fences (∼34 nm radius, ∼400 ms lifetime). Dynamic clustering, trapping and immobilization also occur in larger nanocluster zones (120-180 nm radius) with longer lifetimes (11 ± 1 s), in a strongly cholesterol-sensitive manner. Cholesterol depletion increases the size and average duration of the clustering phenomenon; cholesterol enrichment has the opposite effect. The disclosed high proportion of mAb35-crosslinked immobile receptors, together with their anomalous, cholesterol-sensitive diffusion and clustering, provides new insights into the antibody-enhanced antigenic modulation that leads to physiopathological internalization and degradation of receptors in myasthenia.A preliminary version of this work has appeared in the biorXiv repository: https://www.biorxiv.org/content/10.1101/744664v1. The study was not pre-registered.

Single-molecule mRNA and translation imaging in neurons

2021 ◽  
Author(s):  
Jessica Mitchell ◽  
Jeffrey A. Chao
Keyword(s):  
Single Molecule ◽  
Neuronal Plasticity ◽  
Behavioural Change ◽  
Live Cells ◽  
Local Translation ◽  
Single Molecule Fluorescence ◽  
Technical Developments ◽  
Mrna Transcripts ◽  
Neuronal Compartments

Memory-relevant neuronal plasticity is believed to require local translation of new proteins at synapses. Understanding this process has necessitated the development of tools to visualize mRNA within relevant neuronal compartments. In this review, we summarize the technical developments that now enable mRNA transcripts and their translation to be visualized at single-molecule resolution in both fixed and live cells. These tools include single-molecule fluorescence in situ hybridization (smFISH) to visualize mRNA in fixed cells, MS2/PP7 labelling for live mRNA imaging and SunTag labelling to observe the emergence of nascent polypeptides from a single translating mRNA. The application of these tools in cultured neurons and more recently in whole brains promises to revolutionize our understanding of local translation in the neuronal plasticity that underlies behavioural change.

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