A simple method defines 3D morphology and axon projections of filled neurons in a small CNS volume: Steps toward understanding functional network circuitry
| Autor: | Mike Hull, Deborah Conte, Alan Roberts, Roman Borisyuk |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Neurons
0301 basic medicine Nervous system Patch-Clamp Techniques Computer science General Neuroscience 3D reconstruction Network mapping Axons 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine.anatomical_structure Simple (abstract algebra) Larva Biological neural network medicine Animals Neuron Axon Neuroscience 030217 neurology & neurosurgery Volume (compression) |
| Zdroj: | Journal of Neuroscience Methods. 351:109062 |
| ISSN: | 0165-0270 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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