scholarly journals A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits

2018 ◽  
Author(s):  
Jonathan T. Ting ◽  
Brian Kalmbach ◽  
Peter Chong ◽  
Rebecca de Frates ◽  
C. Dirk Keene ◽  
...  
Keyword(s):  
Human Brain ◽  
Genetic Manipulation ◽  
Ex Vivo ◽  
Molecular Genetic ◽  
Brain Slices ◽  
Cell Types ◽  
Brain Cell ◽  
Membrane Properties ◽  
Genetic Tools ◽  
Adult Human

AbstractThe powerful suite of available genetic tools is driving tremendous progress in understanding mouse brain cell types and circuits. However, the degree of conservation in human remains largely unknown in large part due to the lack of such tools and healthy tissue preparations. To close this gap, we describe a robust and stable adult human neurosurgically-derived ex vivo acute and cultured neocortical brain slice system optimized for rapid molecular-genetic manipulation. Surprisingly, acute human brain slices exhibited exceptional viability, and neuronal intrinsic membrane properties could be assayed for at least three days. Maintaining adult human slices in culture under sterile conditions further enabled the application of viral tools to drive rapid expression of exogenous transgenes. Widespread neuron-specific labeling was achieved as early as two days post infection with HSV-1 vectors, with virally-transduced neurons exhibiting membrane properties largely comparable to uninfected neurons over this short timeframe. Finally, we demonstrate the suitability of this culture paradigm for optical manipulation and monitoring of neuronal activity using genetically encoded probes, opening a path for applying modern molecular-genetic tools to study human brain circuit function.

scholarly journals A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Jonathan T. Ting ◽  
Brian Kalmbach ◽  
Peter Chong ◽  
Rebecca de Frates ◽  
C. Dirk Keene ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Molecular Genetic ◽  
Cell Types ◽  
Genetic Dissection ◽  
Experimental Platform ◽  
Adult Human

Demonstration of JC virus by immunofluorescence in multiple cell types in experimentally infected adult human brain cell cultures

1982 ◽  
Vol 54 (2) ◽  
pp. 189-196 ◽  
Author(s):  
Zofia Wroblewska ◽  
Peter G.E. Kennedy ◽  
Mary C. Wellish ◽  
Robert P. Lisak ◽  
Donald H. Gilden
Keyword(s):  
Human Brain ◽  
Cell Cultures ◽  
Jc Virus ◽  
Cell Types ◽  
Brain Cell ◽  
Adult Human Brain ◽  
Adult Human ◽  
Infected Adult ◽  
Multiple Cell ◽  
Brain Cell Cultures

scholarly journals Multicellular ‘hotspots’ harbour high-grade potential in lower-grade gliomas

2021 ◽  
Author(s):  
Alastair J Kirby ◽  
José P Lavrador ◽  
Istvan Bodi ◽  
Francesco Vergani ◽  
Ranjeev Bhangoo ◽  
...  
Keyword(s):  
Human Brain ◽  
Brain Tissue ◽  
Aminolevulinic Acid ◽  
Ex Vivo ◽  
Brain Slices ◽  
Glioma Cells ◽  
Malignant Progression ◽  
Lower Grade ◽  
High Grade ◽  
Human Brain Tissue

Abstract Background Lower-grade gliomas may be indolent for many years before developing malignant behaviour. The reasons mechanisms underlying malignant progression remain unclear. Methods We collected blocks of live human brain tissue donated by people undergoing glioma resection. The tissue blocks extended through the peritumoral cortex and into the glioma. The living human brain tissue was cut into ex vivo brain slices and bathed in 5-aminolevulinic acid (5-ALA). High-grade glioma cells avidly take up 5-aminolevulinic acid (5-ALA) and accumulate high levels of the fluorescent metabolite, Protoporphyrin IX (PpIX). We exploited the PpIX fluorescence emitted by higher-grade glioma cells to investigate the earliest stages of malignant progression in lower-grade gliomas. Results We found sparsely-distributed ‘hot-spots’ of PpIX-positive cells in living lower-grade glioma tissue. Glioma cells and endothelial cells formed part of the PpIX hotspots. Glioma cells in PpIX hotspots were IDH1 mutant and expressed nestin suggesting they had acquired stem-like properties. Spatial analysis with 5-ALA conjugated quantum dots indicated that these glioma cells replicated adjacent to blood vessels. PpIX hotspots formed in the absence of angiogenesis. Conclusion Our data show that PpIX hotspots represent microdomains of cells with high-grade potential within lower-grade gliomas and identify locations where malignant progression could start.

scholarly journals Amyloid-β Oligomers Induce Differential Gene Expression in Adult Human Brain Slices

2012 ◽  
Vol 287 (10) ◽  
pp. 7436-7445 ◽  
Author(s):  
Adriano Sebollela ◽  
Leo Freitas-Correa ◽  
Fabio F. Oliveira ◽  
Andrea C. Paula-Lima ◽  
Leonardo M. Saraiva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Human Brain ◽  
Differential Gene Expression ◽  
Brain Slices ◽  
Amyloid Β ◽  
Adult Human Brain ◽  
Adult Human ◽  
Differential Gene

Localization of migraine susceptibility genes in human brain by single-cell RNA sequencing

Cephalalgia ◽  
2018 ◽  
Vol 38 (13) ◽  
pp. 1976-1983 ◽  
Author(s):  
William Renthal
Keyword(s):  
Human Brain ◽  
Single Cell ◽  
Rna Sequencing ◽  
Expression Profiles ◽  
Cell Types ◽  
Susceptibility Genes ◽  
Brain Cell ◽  
Cell Type ◽  
Single Cell Rna Sequencing ◽  
Brain Cell Types

Background Migraine is a debilitating disorder characterized by severe headaches and associated neurological symptoms. A key challenge to understanding migraine has been the cellular complexity of the human brain and the multiple cell types implicated in its pathophysiology. The present study leverages recent advances in single-cell transcriptomics to localize the specific human brain cell types in which putative migraine susceptibility genes are expressed. Methods The cell-type specific expression of both familial and common migraine-associated genes was determined bioinformatically using data from 2,039 individual human brain cells across two published single-cell RNA sequencing datasets. Enrichment of migraine-associated genes was determined for each brain cell type. Results Analysis of single-brain cell RNA sequencing data from five major subtypes of cells in the human cortex (neurons, oligodendrocytes, astrocytes, microglia, and endothelial cells) indicates that over 40% of known migraine-associated genes are enriched in the expression profiles of a specific brain cell type. Further analysis of neuronal migraine-associated genes demonstrated that approximately 70% were significantly enriched in inhibitory neurons and 30% in excitatory neurons. Conclusions This study takes the next step in understanding the human brain cell types in which putative migraine susceptibility genes are expressed. Both familial and common migraine may arise from dysfunction of discrete cell types within the neurovascular unit, and localization of the affected cell type(s) in an individual patient may provide insight into to their susceptibility to migraine.

Physiological Properties of Central Medial and Central Lateral Amygdala Neurons

1999 ◽  
Vol 82 (4) ◽  
pp. 1843-1854 ◽  
Author(s):  
Marzia Martina ◽  
Sébastien Royer ◽  
Denis Paré
Keyword(s):  
Conditioned Fear ◽  
Brain Slices ◽  
Cell Types ◽  
Membrane Depolarization ◽  
Membrane Properties ◽  
Repetitive Firing ◽  
Inward Rectification ◽  
Current Pulses ◽  
Outward Rectification ◽  
Physiological Properties

Mounting evidence implicates the central (CE) nucleus of the amygdala in the mediation of classically conditioned fear responses. However, little data are available regarding the intrinsic membrane properties of CE amygdala neurons. Here, we characterized the physiological properties of CE medial (CEM) and CE lateral (CEL) amygdala neurons using whole cell recordings in brain slices maintained in vitro. Several classes of CE neurons were distinguished on the basis of their physiological properties. Most CEM cells (95%), here termed “late-firing neurons,” displayed a marked voltage- and time-dependent outward rectification in the depolarizing direction. This phenomenon was associated with a conspicuous delay between the onset of depolarizing current pulses and the first action potential. During this delay, the membrane potential ( V m) depolarized slowly, the steepness of this depolarizing ramp increasing as the prepulse V m was hyperpolarized from −60 to −90 mV. Low extracellular concentrations of 4-aminopyridine (30 μM) reversibly abolished the outward rectification and the delay to firing. Late-firing CEM neurons displayed a continuum of repetitive firing properties with cells generating single spikes at one pole and high-frequency (≥90 Hz) spike bursts at the other. In contrast, only 56% of CEL cells displayed the late-firing behavior prevalent among CEM neurons. Moreover, these CEL neurons only generated single spikes in response to membrane depolarization. A second major class of CEL cells (38%) lacked the characteristic delay to firing observed in CEM cells, generated single spikes in response to membrane depolarization, and displayed various degrees of inward rectification in the hyperpolarizing direction. In both regions of the CE nucleus, two additional cell types were encountered infrequently (≤ 6% of our samples). One type of neurons, termed “low-threshold bursting cells” had a behavior reminiscent of thalamocortical neurons. The second type of cells, called “fast-spiking cells,” generated brief action potentials at high rates with little spike frequency adaptation in response to depolarizing current pulses. These findings indicate that the CE nucleus contains several types of neurons endowed with distinct physiological properties. Moreover, these various cell types are not distributed uniformly in the medial and lateral sector of the CE nucleus. This heterogeneity parallels anatomic data indicating that these subnuclei are part of different circuits.

Differential expression of peroxiredoxin subtypes in human brain cell types

1999 ◽  
Vol 56 (2) ◽  
pp. 206-212
Author(s):  
Theodore A. Sarafian ◽  
M. Anthony Verity ◽  
Harry V. Vinters ◽  
Charles C.-Y. Shih ◽  
Liangru Shi ◽  
...  
Keyword(s):  
Human Brain ◽  
Differential Expression ◽  
Cell Types ◽  
Brain Cell ◽  
Brain Cell Types

scholarly journals Comparison Between Human and Rodent Neurons for Persistent Activity Performance: A Biologically Plausible Computational Investigation

2021 ◽  
Vol 15 ◽  
Author(s):  
Qian Zhang ◽  
Yi Zeng ◽  
Tielin Zhang ◽  
Taoyi Yang
Keyword(s):  
Human Brain ◽  
Computational Models ◽  
Brain Cell ◽  
Membrane Properties ◽  
Accurate Information ◽  
Computational Investigation ◽  
Multi Scale ◽  
Neocortical Neurons ◽  
Activity Performance ◽  
Shed Light

Elucidating the multi-scale detailed differences between the human brain and other brains will help shed light on what makes us unique as a species. Computational models help link biochemical and anatomical properties to cognitive functions and predict key properties of the cortex. Here, we present a detailed human neocortex network, with all human neuron parameters derived from the newest Allen Brain human brain cell database. Compared with that of rodents, the human neural network maintains more complete and accurate information under the same graphic input. Unique membrane properties in human neocortical neurons enhance the human brain’s capacity for signal processing.

Cell Type–Specific Nuclei Markers: The Need for Human Brain Research to Go Nuclear

2021 ◽  
pp. 107385842110373
Author(s):  
James A. Wiseman ◽  
Mike Dragunow ◽  
Thomas I.-H. Park
Keyword(s):  
Human Brain ◽  
Brain Research ◽  
Cell Types ◽  
Brain Cell ◽  
Normal Brain ◽  
Cell Type ◽  
Human Brain Tissue ◽  
Nuclear Markers ◽  
Cell Type Specific ◽  
Immunohistochemical Studies

Identifying and interrogating cell type–specific populations within the heterogeneous milieu of the human brain is paramount to resolving the processes of normal brain homeostasis and the pathogenesis of neurological disorders. While brain cell type–specific markers are well established, most are localized on cellular membranes or within the cytoplasm, with limited literature describing those found in the nucleus. Due to the complex cytoarchitecture of the human brain, immunohistochemical studies require well-defined cell-specific nuclear markers for more precise and efficient quantification of the cellular populations. Furthermore, efficient nuclear markers are required for cell type–specific purification and transcriptomic interrogation of archived human brain tissue through nuclei isolation–based RNA sequencing. To sate the growing demand for robust cell type–specific nuclear markers, we thought it prudent to comprehensively review the current literature to identify and consolidate a novel series of robust cell type–specific nuclear markers that can assist researchers across a range of neuroscientific disciplines. The following review article collates and discusses several key and prospective cell type–specific nuclei markers for each of the major human brain cell types; it then concludes by discussing the potential applications of cell type–specific nuclear workflows and the power of nuclear-based neuroscientific research.

scholarly journals Cellular and genetic drivers of RNA editing variation in the human brain

2021 ◽  
Author(s):  
Ryn Cuddleston ◽  
Junhao Li ◽  
Xuanjia Fan ◽  
Alexey Kozenkov ◽  
Matthew Lalli ◽  
...  
Keyword(s):  
Human Brain ◽  
Rna Sequencing ◽  
Rna Editing ◽  
Genetic Regulation ◽  
Cell Types ◽  
Brain Regions ◽  
Brain Cell ◽  
Cell Type ◽  
Sequencing Data ◽  
Regulatory Effects

Posttranscriptional adenosine-to-inosine modifications amplify the functionality of RNA molecules in the brain, yet the cellular and genetic regulation of RNA editing is poorly described. We quantified base-specific RNA editing across three major cell populations from the human prefrontal cortex: glutamatergic neurons, medial ganglionic eminence GABAergic neurons, and oligodendrocytes. We found more selective editing and RNA hyper-editing in neurons relative to oligodendrocytes. The pattern of RNA editing was highly cell type-specific, with 189,229 cell type-associated sites. The cellular specificity for thousands of sites was confirmed by single nucleus RNA-sequencing. Importantly, cell type-associated sites were enriched in GTEx RNA-sequencing data, edited ~twentyfold higher than all other sites, and variation in RNA editing was predominantly explained by neuronal proportions in bulk brain tissue. Finally, we discovered 661,791 cis-editing quantitative trait loci across thirteen brain regions, including hundreds with cell type-associated features. These data reveal an expansive repertoire of highly regulated RNA editing sites across human brain cell types and provide a resolved atlas linking cell types to editing variation and genetic regulatory effects.

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