A simple method defines 3D morphology and axon projections of filled neurons in a small CNS volume : Steps toward understanding functional network circuitry
Copyright © 2021 Elsevier B.V. All rights reserved..
BACKGROUND: Fundamental to understanding neuronal network function is defining neuron morphology, location, properties, and synaptic connectivity in the nervous system. A significant challenge is to reconstruct individual neuron morphology and connections at a whole CNS scale and bring together functional and anatomical data to understand the whole network.
NEW METHOD: We used a PC controlled micropositioner to hold a fixed whole mount of Xenopus tadpole CNS and replace the stage on a standard microscope. This allowed direct recording in 3D coordinates of features and axon projections of one or two neurons dye-filled during whole-cell recording to study synaptic connections. Neuron reconstructions were normalised relative to the ventral longitudinal axis of the nervous system. Coordinate data were stored as simple text files.
RESULTS: Reconstructions were at 1 μm resolution, capturing axon lengths in mm. The output files were converted to SWC format and visualised in 3D reconstruction software NeuRomantic. Coordinate data are tractable, allowing correction for histological artefacts. Through normalisation across multiple specimens we could infer features of network connectivity of mapped neurons of different types.
COMPARISON WITH EXISTING METHODS: Unlike other methods using fluorescent markers and utilising large-scale imaging, our method allows direct acquisition of 3D data on neurons whose properties and synaptic connections have been studied using whole-cell recording.
CONCLUSIONS: This method can be used to reconstruct neuron 3D morphology and follow axon projections in the CNS. After normalisation to a common CNS framework, inferences on network connectivity at a whole nervous system scale contribute to network modelling to understand CNS function.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:351 |
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| Enthalten in: |
Journal of neuroscience methods - 351(2021) vom: 01. März, Seite 109062 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Conte, Deborah [VerfasserIn] |
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| Links: |
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| Themen: |
Brightfield microscopy |
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| Anmerkungen: |
Date Completed 30.06.2021 Date Revised 06.10.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jneumeth.2020.109062 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM319471063 |
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| 100 | 1 | |a Conte, Deborah |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A simple method defines 3D morphology and axon projections of filled neurons in a small CNS volume |b Steps toward understanding functional network circuitry |
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| 500 | |a Date Completed 30.06.2021 | ||
| 500 | |a Date Revised 06.10.2021 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2021 Elsevier B.V. All rights reserved. | ||
| 520 | |a BACKGROUND: Fundamental to understanding neuronal network function is defining neuron morphology, location, properties, and synaptic connectivity in the nervous system. A significant challenge is to reconstruct individual neuron morphology and connections at a whole CNS scale and bring together functional and anatomical data to understand the whole network | ||
| 520 | |a NEW METHOD: We used a PC controlled micropositioner to hold a fixed whole mount of Xenopus tadpole CNS and replace the stage on a standard microscope. This allowed direct recording in 3D coordinates of features and axon projections of one or two neurons dye-filled during whole-cell recording to study synaptic connections. Neuron reconstructions were normalised relative to the ventral longitudinal axis of the nervous system. Coordinate data were stored as simple text files | ||
| 520 | |a RESULTS: Reconstructions were at 1 μm resolution, capturing axon lengths in mm. The output files were converted to SWC format and visualised in 3D reconstruction software NeuRomantic. Coordinate data are tractable, allowing correction for histological artefacts. Through normalisation across multiple specimens we could infer features of network connectivity of mapped neurons of different types | ||
| 520 | |a COMPARISON WITH EXISTING METHODS: Unlike other methods using fluorescent markers and utilising large-scale imaging, our method allows direct acquisition of 3D data on neurons whose properties and synaptic connections have been studied using whole-cell recording | ||
| 520 | |a CONCLUSIONS: This method can be used to reconstruct neuron 3D morphology and follow axon projections in the CNS. After normalisation to a common CNS framework, inferences on network connectivity at a whole nervous system scale contribute to network modelling to understand CNS function | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Brightfield microscopy | |
| 650 | 4 | |a Functional anatomy | |
| 650 | 4 | |a Multiscale | |
| 650 | 4 | |a Network mapping | |
| 650 | 4 | |a Neuron reconstruction | |
| 650 | 4 | |a SWC | |
| 700 | 1 | |a Borisyuk, Roman |e verfasserin |4 aut | |
| 700 | 1 | |a Hull, Mike |e verfasserin |4 aut | |
| 700 | 1 | |a Roberts, Alan |e verfasserin |4 aut | |
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