In vivo imaging of neural circuit formation in the neonatal mouse barrel cortex

2020 ◽  
Vol 62 (7-8) ◽  
pp. 476-486
Author(s):  
Takuji Iwasato
Keyword(s):  
In Vivo Imaging ◽  
Neural Circuit ◽  
Barrel Cortex ◽  
Neonatal Mouse ◽  
Circuit Formation

scholarly journals In Vivo Imaging of Axonal and Dendritic Structures in Neonatal Mouse Cortex

2013 ◽  
Vol 2014 (1) ◽  
pp. pdb.prot080150 ◽  
Author(s):  
Alberto Cruz-Martin ◽  
Carlos Portera-Cailliau
Keyword(s):  
In Vivo Imaging ◽  
Neonatal Mouse ◽  
Mouse Cortex ◽  
Dendritic Structures

scholarly journals Selective and constructive mechanisms contribute to neural circuit formation in the barrel cortex of the developing rat

2013 ◽  
Vol 04 (07) ◽  
pp. 785-797 ◽  
Author(s):  
Eileen Uribe-Querol ◽  
Eduardo Martínez-Martínez ◽  
Luis Rodrigo Hernández ◽  
Patricia Padilla Cortés ◽  
Horacio Merchant-Larios ◽  
...  
Keyword(s):  
Neural Circuit ◽  
Barrel Cortex ◽  
Circuit Formation ◽  
Developing Rat

scholarly journals All-optical interrogation of neural circuits in behaving mice

2021 ◽  
Author(s):  
Lloyd E. Russell ◽  
Henry W.P. Dalgleish ◽  
Rebecca Nutbrown ◽  
Oliver Gauld ◽  
Dustin Herrmann ◽  
...  
Keyword(s):  
Neural Circuits ◽  
Neural Circuit ◽  
Barrel Cortex ◽  
Brain Area ◽  
Imaging Data ◽  
Temporal Precision ◽  
Two Photon ◽  
All Optical ◽  
The Impact

Recent advances combining two-photon calcium imaging and two-photon optogenetics with digital holography now allow us to read and write neural activity in vivo at cellular resolution with millisecond temporal precision. Such 'all-optical' techniques enable experimenters to probe the impact of functionally defined neurons on neural circuit function and behavioural output with new levels of precision. This protocol describes the experimental strategy and workflow for successful completion of typical all-optical interrogation experiments in awake, behaving head-fixed mice. We describe modular procedures for the setup and calibration of an all-optical system, the preparation of an indicator and opsin-expressing and task-performing animal, the characterization of functional and photostimulation responses and the design and implementation of an all-optical experiment. We discuss optimizations for efficiently selecting and targeting neuronal ensembles for photostimulation sequences, as well as generating photostimulation response maps from the imaging data that can be used to examine the impact of photostimulation on the local circuit. We demonstrate the utility of this strategy using all-optical experiments in three different brain areas - barrel cortex, visual cortex and hippocampus - using different experimental setups. This approach can in principle be adapted to any brain area for all-optical interrogation experiments to probe functional connectivity in neural circuits and for investigating the relationship between neural circuit activity and behaviour.

scholarly journals All-optical electrophysiology reveals excitation, inhibition, and neuromodulation in cortical layer 1

2019 ◽  
Author(s):  
Linlin Z. Fan ◽  
Simon Kheifets ◽  
Urs L. Böhm ◽  
Kiryl D. Piatkevich ◽  
Hao Wu ◽  
...  
Keyword(s):  
Neural Circuit ◽  
Barrel Cortex ◽  
Cortical Layer ◽  
Optical Recording ◽  
Tight Coupling ◽  
Basic Principles ◽  
All Optical ◽  
The Stability ◽  
Circuit Function

AbstractThe stability of neural dynamics arises through a tight coupling of excitatory (E) and inhibitory (I) signals. Genetically encoded voltage indicators (GEVIs) can report both spikes and subthreshold dynamics in vivo, but voltage only reveals the combined effects of E and I synaptic inputs, not their separate contributions individually. Here we combine optical recording of membrane voltage with simultaneous optogenetic manipulation to probe E and I individually in barrel cortex Layer 1 (L1) neurons in awake mice. Our studies reveal how the L1 microcircuit integrates thalamocortical excitation, lateral inhibition and top-down neuromodulatory inputs. We develop a simple computational model of the L1 microcircuit which captures the main features of our data. Together, these results suggest a model for computation in L1 interneurons consistent with their hypothesized role in attentional gating of the underlying cortex. Our results demonstrate that all-optical electrophysiology can reveal basic principles of neural circuit function in vivo.One Sentence SummaryAll-optical electrophysiology revealed the function in awake mice of an inhibitory microcircuit in barrel cortex Layer 1.

scholarly journals Axon guidance at the midline – a live imaging perspective

2020 ◽  
Author(s):  
Alexandre Dumoulin ◽  
Nikole R. Zuñiga ◽  
Esther T. Stoeckli
Keyword(s):  
Molecular Mechanisms ◽  
Neural Circuit ◽  
Surface Expression ◽  
Growth Cones ◽  
Live Imaging ◽  
Growth Speed ◽  
Intermediate Target ◽  
Commissural Axons ◽  
Circuit Formation

ABSTRACTDuring neural circuit formation, axons navigate several choice points to reach their final target. At each one of these intermediate targets, growth cones need to switch responsiveness from attraction to repulsion in order to move on. Molecular mechanisms that allow for the precise timing of surface expression of a new set of receptors that support the switch in responsiveness are difficult to study in vivo. Mostly, mechanisms are inferred from the observation of snapshots of many different growth cones analyzed in different preparations of tissue harvested at distinct time points. However, to really understand the behavior of growth cones at choice points, a single growth cone should be followed arriving at and leaving the intermediate target.Here, we describe a spinal cord preparation that allows for live imaging of individual axons during navigation in their intact environment. The possibility to observe single growth cones navigating their intermediate target allows for measuring growth speed, changes in morphology, or aberrant behavior. Moreover, observation of the intermediate target – the floor plate – revealed its active participation and interaction with commissural axons during midline crossing.Summary statementLive tracking of single growth cones is more informative about axonal behavior during navigation than inference of behavior from the analyses of snapshots of different growth cones.

scholarly journals Architects in neural circuit design: Glia control neuron numbers and connectivity

2013 ◽  
Vol 203 (3) ◽  
pp. 395-405 ◽  
Author(s):  
Megan M. Corty ◽  
Marc R. Freeman
Keyword(s):  
Nervous System ◽  
Circuit Design ◽  
Glial Cells ◽  
Neural Development ◽  
Molecular Mechanisms ◽  
Neural Circuit ◽  
Neuronal Circuit ◽  
Circuit Formation

Glia serve many important functions in the mature nervous system. In addition, these diverse cells have emerged as essential participants in nearly all aspects of neural development. Improved techniques to study neurons in the absence of glia, and to visualize and manipulate glia in vivo, have greatly expanded our knowledge of glial biology and neuron–glia interactions during development. Exciting studies in the last decade have begun to identify the cellular and molecular mechanisms by which glia exert control over neuronal circuit formation. Recent findings illustrate the importance of glial cells in shaping the nervous system by controlling the number and connectivity of neurons.

Fluorescent proteins for in vivo imaging, where's the biliverdin?

2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez
Keyword(s):  
Hydrogen Peroxide ◽  
In Vivo Imaging ◽  
Near Infrared ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Imaging ◽  
Infrared Light ◽  
Red Fluorescent Protein ◽  
Color Palette

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.

In vivo imaging of beta-amyloid load in transgenic rodents with [F-18]FDDNP microPET

2005 ◽  
Vol 25 (1_suppl) ◽  
pp. S588-S588
Author(s):  
Vladimir Kepe ◽  
Gregory M Cole ◽  
Jie Liu ◽  
Dorothy G Flood ◽  
Stephen P Trusko ◽  
...  
Keyword(s):  
In Vivo Imaging ◽  
Beta Amyloid ◽  
Amyloid Load

In vivo imaging of liver injury and regeneration by functional two-photon microscopy

2016 ◽  
Vol 54 (12) ◽  
pp. 1343-1404
Author(s):  
A Ghallab ◽  
R Reif ◽  
R Hassan ◽  
AS Seddek ◽  
JG Hengstler
Keyword(s):  
Liver Injury ◽  
In Vivo Imaging ◽  
Two Photon Microscopy ◽  
Two Photon
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