scholarly journals In Vivo Functional Mapping of a Cortical Column at Single-Neuron Resolution

Cell Reports ◽  
2019 ◽  
Vol 27 (5) ◽  
pp. 1319-1326.e5 ◽  
Author(s):  
Carsten H. Tischbirek ◽  
Takahiro Noda ◽  
Manabu Tohmi ◽  
Antje Birkner ◽  
Israel Nelken ◽  
...  
Keyword(s):  
Single Neuron ◽  
Functional Mapping ◽  
Cortical Column

scholarly journals A method for single-neuron chronic recording from the retina in awake mice

Science ◽  
2018 ◽  
Vol 360 (6396) ◽  
pp. 1447-1451 ◽  
Author(s):  
Guosong Hong ◽  
Tian-Ming Fu ◽  
Mu Qiao ◽  
Robert D. Viveros ◽  
Xiao Yang ◽  
...  
Keyword(s):  
Retinal Ganglion Cells ◽  
Visual Information ◽  
Single Neuron ◽  
Ganglion Cells ◽  
Ex Vivo ◽  
Neural Circuitry ◽  
Retinal Ganglion ◽  
Chronic Recording ◽  
The Brain

The retina, which processes visual information and sends it to the brain, is an excellent model for studying neural circuitry. It has been probed extensively ex vivo but has been refractory to chronic in vivo electrophysiology. We report a nonsurgical method to achieve chronically stable in vivo recordings from single retinal ganglion cells (RGCs) in awake mice. We developed a noncoaxial intravitreal injection scheme in which injected mesh electronics unrolls inside the eye and conformally coats the highly curved retina without compromising normal eye functions. The method allows 16-channel recordings from multiple types of RGCs with stable responses to visual stimuli for at least 2 weeks, and reveals circadian rhythms in RGC responses over multiple day/night cycles.

scholarly journals Combining single-cell RNA-sequencing with a molecular atlas unveils new markers for Caenorhabditis elegans neuron classes

2020 ◽  
Author(s):  
Ramiro Lorenzo ◽  
Michiho Onizuka ◽  
Matthieu Defrance ◽  
Patrick Laurent
Keyword(s):  
Caenorhabditis Elegans ◽  
Single Cell ◽  
Rna Sequencing ◽  
Cell Fate ◽  
Single Neuron ◽  
Genetic Manipulations ◽  
C Elegans ◽  
Single Cell Rna Sequencing ◽  
Normal Differentiation

Abstract Single-cell RNA-sequencing (scRNA-seq) of the Caenorhabditis elegans nervous system offers the unique opportunity to obtain a partial expression profile for each neuron within a known connectome. Building on recent scRNA-seq data and on a molecular atlas describing the expression pattern of ∼800 genes at the single cell resolution, we designed an iterative clustering analysis aiming to match each cell-cluster to the ∼100 anatomically defined neuron classes of C. elegans. This heuristic approach successfully assigned 97 of the 118 neuron classes to a cluster. Sixty two clusters were assigned to a single neuron class and 15 clusters grouped neuron classes sharing close molecular signatures. Pseudotime analysis revealed a maturation process occurring in some neurons (e.g. PDA) during the L2 stage. Based on the molecular profiles of all identified neurons, we predicted cell fate regulators and experimentally validated unc-86 for the normal differentiation of RMG neurons. Furthermore, we observed that different classes of genes functionally diversify sensory neurons, interneurons and motorneurons. Finally, we designed 15 new neuron class-specific promoters validated in vivo. Amongst them, 10 represent the only specific promoter reported to this day, expanding the list of neurons amenable to genetic manipulations.

scholarly journals Dissection of Cortical Microcircuits by Single-Neuron Stimulation In Vivo

2012 ◽  
Vol 22 (16) ◽  
pp. 1459-1467 ◽  
Author(s):  
Alex C. Kwan ◽  
Yang Dan
Keyword(s):  
Single Neuron ◽  
Cortical Microcircuits

Long-lasting single-neuron labeling by in vivo electroporation without microscopic guidance

2013 ◽  
Vol 218 (2) ◽  
pp. 139-147 ◽  
Author(s):  
Kei Oyama ◽  
Shinya Ohara ◽  
Sho Sato ◽  
Fuyuki Karube ◽  
Fumino Fujiyama ◽  
...  
Keyword(s):  
Single Neuron ◽  
In Vivo Electroporation

scholarly journals In vivo functional mapping of the conserved protein domains within murine Themis1

2014 ◽  
Vol 92 (8) ◽  
pp. 721-728 ◽  
Author(s):  
Ekaterina Zvezdova ◽  
Jan Lee ◽  
Dalal El‐Khoury ◽  
Valarie Barr ◽  
Itoro Akpan ◽  
...  
Keyword(s):  
Protein Domains ◽  
Functional Mapping

Safety of Hybrid Electrodes for Single-Neuron Recordings in Humans

Neurosurgery ◽  
2013 ◽  
Vol 73 (1) ◽  
pp. 78-85 ◽  
Author(s):  
Stefan Hefft ◽  
Armin Brandt ◽  
Stefan Zwick ◽  
Dominik von Elverfeldt ◽  
Irina Mader ◽  
...  
Keyword(s):  
Single Neuron ◽  
Electrode Implantation ◽  
Intraoperative Computed Tomography ◽  
Postoperative Mri ◽  
Depth Electrodes ◽  
Magnetic Resonance Imaging Mri ◽  
Clinical Mri

Abstract BACKGROUND: Intracranial in vivo recordings of individual neurons in humans are increasingly performed for a better understanding of the mechanisms of epileptogenesis and of the neurobiological basis of cognition. So far, information about the safety of stereotactic implantations and of magnetic resonance imaging (MRI) with hybrid depth electrodes is scarce. OBJECTIVE: The aim of this study was to assess neurosurgical safety of implantations, recordings, and imaging using hybrid electrodes in humans. METHODS: Perioperative and long-term safety of implantation of a total of 88 hybrid depth electrodes with integrated microwires was assessed retrospectively in 25 consecutive epilepsy patients who underwent implantation of electrodes from 2007 to 2011 based on electronically stored charts. Safety aspects of MRI are reported from both in vitro and in vivo investigations. Precision of electrode implantation is evaluated based on intraoperative computed tomography and pre- and postoperative MRI. RESULTS: There was no clinically relevant morbidity associated with the use of hybrid electrodes in any of the patients. Precision of recordings from the targets aimed at was similar to that of standard depth electrodes. In vitro studies demonstrated the absence of relevant heating of hybrid electrodes with newly designed connectors with MRI at 1.5 T, corresponding to well-tolerated clinical MRI in patients. CONCLUSION: Given the technical approach described here, precise targeting and safe use are possible with hybrid electrodes containing microwires for in vivo recording of human neuronal units.

scholarly journals Functional Mapping of Adhesiveness on Live Cells Reveals How Guidance Phenotypes Can Emerge From Complex Spatiotemporal Integrin Regulation

2021 ◽  
Vol 9 ◽  
Author(s):  
Philippe Robert ◽  
Martine Biarnes-Pelicot ◽  
Nicolas Garcia-Seyda ◽  
Petra Hatoum ◽  
Dominique Touchard ◽  
...  
Keyword(s):  
Distance Effect ◽  
Cell Polarization ◽  
Functional Mapping ◽  
Live Cells ◽  
Long Distance ◽  
Adhesion Properties ◽  
Functional Assays ◽  
New Perspective

Immune cells have the ubiquitous capability to migrate disregarding the adhesion properties of the environment, which requires a versatile adaptation of their adhesiveness mediated by integrins, a family of specialized adhesion proteins. Each subtype of integrins has several ligands and several affinity states controlled by internal and external stimuli. However, probing cell adhesion properties on live cells without perturbing cell motility is highly challenging, especially in vivo. Here, we developed a novel in vitro method using micron-size beads pulled by flow to functionally probe the local surface adhesiveness of live and motile cells. This method allowed a functional mapping of the adhesiveness mediated by VLA-4 and LFA-1 integrins on the trailing and leading edges of live human T lymphocytes. We show that cell polarization processes enhance integrin-mediated adhesiveness toward cell rear for VLA-4 and cell front for LFA-1. Furthermore, an inhibiting crosstalk of LFA-1 toward VLA-4 and an activating crosstalk of VLA-4 toward LFA-1 were found to modulate cell adhesiveness with a long-distance effect across the cell. These combined signaling processes directly support the bistable model that explains the emergence of the versatile guidance of lymphocyte under flow. Molecularly, Sharpin, an LFA-1 inhibitor in lymphocyte uropod, was found involved in the LFA-1 deadhesion of lymphocytes; however, both Sharpin and Myosin inhibition had a rather modest impact on adhesiveness. Quantitative 3D immunostaining identified high-affinity LFA-1 and VLA-4 densities at around 50 and 100 molecules/μm2 in basal adherent zones, respectively. Interestingly, a latent adhesiveness of dorsal zones was not grasped by immunostaining but assessed by direct functional assays with beads. The combination of live functional assays, molecular imaging, and genome editing is instrumental to characterizing the spatiotemporal regulation of integrin-mediated adhesiveness at molecular and cell scales, which opens a new perspective to decipher sophisticated phenotypes of motility and guidance.

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