Holographic Functional Calcium Imaging of Neuronal Circuit Activity

Castañares, Michael Lawrence; Stuart, G. W.; Daria, Vincent Ricardo

doi:10.1007/978-981-10-9020-2_8

Cited by 6 publications

(8 citation statements)

References 67 publications

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“…For patch-clamp recording and 2P imaging, tissue slices were transferred to a custom-built 2P microscope. Whole-cell recordings were made from the soma of L5PNs with 4-6 MΩ pipettes filled with an intracellular solution containing (in mM): 115 K-gluconate, 20 KCl, 10 HEPES, 10 phosphocreatine, 4.0 ATP-Mg, and 0.3 GTP (Castanares et al, 2019). During recording slices were kept at 33±1 O C with the temperature of the extracellular solution regulated by a proportional integral derivative controller (Inkbird ITC-100).…”

Section: Electrophysiologymentioning

confidence: 99%

“…We used a custom-built two-photon microscope as previously reported (Go et al, 2013, Go et al, 2016 and modified it to include a widefield imaging camera for holographic calcium imaging (Castanares et al, 2016, Castanares et al, 2019 (Supplementary Figure 2). The microscope uses a near-infrared femtosecond laser (Chameleon, Coherent Scientific) and galvanometer scanning mirrors to render a 3D image of the patched neuron ( Supplementary Figure 7a).…”

Section: Two-photon Holographic Calcium Imagingmentioning

confidence: 99%

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Dendritic spikes in apical oblique dendrites of cortical layer 5 pyramidal neurons

Castañares

Stuart

Daria

2020

Preprint

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Dendritic spikes in layer 5 pyramidal neurons (L5PNs) play a major role in cortical computation. While dendritic spikes have been studied extensively in apical and basal dendrites of L5PNs, whether oblique dendrites, which ramify in the input layers of the cortex, also generate dendritic spikes is unknown. Here we report the existence of dendritic spikes in apical oblique dendrites of L5PNs. In silico investigations indicate that oblique branch spikes are triggered by brief, low-frequency action potential (AP) trains (~40 Hz) and are characterized by a fast sodium spike followed by activation of voltage-gated calcium channels. In vitro experiments confirmed the existence of oblique branch spikes in L5PNs during brief AP trains at frequencies of around 60 Hz. Oblique branch spikes offer new insights into branch-specific computation in L5PNs and may be critical for sensory processing in the input layers of the cortex.

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Section: Electrophysiologymentioning

confidence: 99%

Section: Two-photon Holographic Calcium Imagingmentioning

confidence: 99%

Dendritic spikes in apical oblique dendrites of cortical layer 5 pyramidal neurons

Castañares

Stuart

Daria

2020

Preprint

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“…Electrophysiological recording at the dendrite where the spike occurs provides for an accurate characterization of D-spikes but could be challenging especially when the dendrite is thinner than the tip of the electrode. Optical recording via calcium or voltage indicators can also be used to record dendritic activity [33]. However, both optical and electrophysiological approaches to record dendritic activity become ineffective after dendrites are severed from the neuron.…”

Section: Multi-compartmental Modelmentioning

confidence: 99%

“…Somatosensory cortical brain slices were prepared using a vibratome and perfused with oxygenated (95% O 2 /5% CO 2 ) extracellular solution containing (in mM): 125 NaCl, 25 NaHCO 3 , 25 glucose, 3 KCl, 1.25 NaH 2 PO 4 , 2 CaCl 2 , 1 MgCl 2 , pH 7.4. Whole-cell patchclamp recordings were made with 4-6 MΩ pipettes filled with an intracellular solution containing (in mM): 115 K-gluconate, 20 KCl, 10 HEPES, 10 phosphocreatine, 4.0 ATP-Mg, 0.3 GTP, and 0.13 Alexa Fluor 488 dye (Sigma Aldrich) [33].…”

Section: Oblique Branch Spike In L5pns In Vitromentioning

confidence: 99%

Analyzing Branch‐specific Dendritic Spikes Using an Ultrafast Laser Scalpel

2020

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Dendritic spikes facilitate neuronal computation and they have been reported to occur in various regions of the dendritic tree of cortical neurons. Spikes that occur only on a select few branches are particularly difficult to analyze especially in complex and intertwined dendritic arborizations where highly localized application of pharmacological blocking agents is not feasible. Here, we present a technique based on highly targeted dendrotomy to tease out and study dendritic spikes that occur in oblique branches of cortical layer five pyramidal neurons. We first analyze the effect of cutting dendrites in silico and then confirmed in vitro using an ultrafast laser scalpel. A dendritic spike evoked in an oblique branch manifests at the soma as an increase in the afterdepolarization (ADP). The spikes are branch-specific since not all but only a few oblique dendrites are observed to evoke spikes. Both our model and experiments show that cutting certain oblique branches, where dendritic spikes are evoked, curtailed the increase in the ADP. On the other hand, cutting neighboring oblique branches that do not evoke spikes maintained the ADP. Our results show that highly targeted dendrotomy can facilitate causal analysis of how branch-specific dendritic spikes influence neuronal output.

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“…Recent advances using in vivo optical methods including calcium imaging have provided a rich source of complex micro anatomical and dynamic neuronal data, including in awake behaving subjects ( Ohki and Reid, 2014 ; Romano et al, 2017 ; Castanares et al, 2019 ). Recent analysis approaches for these data include the method developed by Romano and associates, to analyze neuronal population dynamics ( Romano et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Functional Neuronal Topography: A Statistical Approach to Micro Mapping Neuronal Location

Jacques

Wright

Chaaya

et al. 2018

Front. Neural Circuits

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In order to understand the relationship between neuronal organization and behavior, precise methods that identify and quantify functional cellular ensembles are required. This is especially true in the quest to understand the mechanisms of memory. Brain structures involved in memory formation and storage, as well as the molecular determinates of memory are well-known, however, the microanatomy of functional neuronal networks remain largely unidentified. We developed a novel approach to statistically map molecular markers in neuronal networks through quantitative topographic measurement. Brain nuclei and their subdivisions are well-defined – our approach allows for the identification of new functional micro-regions within established subdivisions. A set of analytic methods relevant for measurement of discrete neuronal data across a diverse range of brain subdivisions are presented. We provide a methodology for the measurement and quantitative comparison of functional micro-neural network activity based on immunohistochemical markers matched across individual brains using micro-binning and heat mapping within brain sub-nuclei. These techniques were applied to the measurement of different memory traces, allowing for greater understanding of the functional encoding within sub-nuclei and its behavior mediated change. These approaches can be used to understand other functional and behavioral questions, including sub-circuit organization, normal memory function and the complexities of pathology. Precise micro-mapping of functional neuronal topography provides essential data to decode network activity underlying behavior.

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Holographic Functional Calcium Imaging of Neuronal Circuit Activity

Cited by 6 publications

References 67 publications

Dendritic spikes in apical oblique dendrites of cortical layer 5 pyramidal neurons

Dendritic spikes in apical oblique dendrites of cortical layer 5 pyramidal neurons

Analyzing Branch‐specific Dendritic Spikes Using an Ultrafast Laser Scalpel

Functional Neuronal Topography: A Statistical Approach to Micro Mapping Neuronal Location

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