scholarly journals Cell type-specific modulation of sensory and affective components of itch in the periaqueductal gray

2018 ◽  
Author(s):  
Vijay K Samineni ◽  
Jose G Grajales-Reyes ◽  
Saranya S Sundaram ◽  
Robert W Gereau
Keyword(s):  
Neural Circuits ◽  
Periaqueductal Gray ◽  
Gabaergic Neurons ◽  
Cell Type ◽  
Glutamatergic Neurons ◽  
Neuronal Populations ◽  
Chronic Itch ◽  
Cell Type Specific ◽  
Bidirectional Modulation ◽  
Affective Components

Itch is a distinct aversive sensation that elicits a strong urge to scratch. Despite recent advances in our understanding of the peripheral basis of itch, we know very little regarding how central neural circuits modulate acute and chronic itch processing. Here we establish the causal contributions of defined periaqueductal gray (PAG) neuronal populations in itch modulation. Chemogenetic manipulations demonstrate bidirectional modulation of scratching by neurons in the PAG. Fiber photometry studies show that activity of GABAergic and glutamatergic neurons in the PAG is modulated in an opposing manner during chloroquine-evoked scratching. Furthermore, activation of PAG GABAergic neurons or inhibition of glutamatergic neurons resulted in attenuation of scratching in both acute and chronic pruritis. Surprisingly, PAG GABAergic neurons, but not glutamatergic neurons, seem to encode the aversive component of itch. Thus, the PAG represents a neuromodulatory hub that regulates both the sensory and affective aspects of acute and chronic itch.

scholarly journals Cell type-specific modulation of sensory and affective components of itch in the periaqueductal gray

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Vijay K. Samineni ◽  
Jose G. Grajales-Reyes ◽  
Saranya S. Sundaram ◽  
Judy J. Yoo ◽  
Robert W. Gereau
Keyword(s):  
Neural Circuits ◽  
Periaqueductal Gray ◽  
Gabaergic Neurons ◽  
Cell Type ◽  
Glutamatergic Neurons ◽  
Neuronal Populations ◽  
Chronic Itch ◽  
Cell Type Specific ◽  
Bidirectional Modulation ◽  
Affective Components

Abstract Itch is a distinct aversive sensation that elicits a strong urge to scratch. Despite recent advances in our understanding of the peripheral basis of itch, we know very little regarding how central neural circuits modulate acute and chronic itch processing. Here we establish the causal contributions of defined periaqueductal gray (PAG) neuronal populations in itch modulation in mice. Chemogenetic manipulations demonstrate bidirectional modulation of scratching by neurons in the PAG. Fiber photometry studies show that activity of GABAergic and glutamatergic neurons in the PAG is modulated in an opposing manner during chloroquine-evoked scratching. Furthermore, activation of PAG GABAergic neurons or inhibition of glutamatergic neurons resulted in attenuation of scratching in both acute and chronic pruritis. Surprisingly, PAG GABAergic neurons, but not glutamatergic neurons, may encode the aversive component of itch. Thus, the PAG represents a neuromodulatory hub that regulates both the sensory and affective aspects of acute and chronic itch.

scholarly journals Fast and reversible neural inactivation in macaque cortex by optogenetic stimulation of GABAergic neurons

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Abhishek De ◽  
Yasmine El-Shamayleh ◽  
Gregory D Horwitz
Keyword(s):  
Neural Activity ◽  
Neural Circuits ◽  
Gabaergic Neurons ◽  
Visual Sensitivity ◽  
Cell Type ◽  
Behavioral Deficits ◽  
Inhibitory Cell ◽  
Optogenetic Stimulation ◽  
Cell Type Specific ◽  
Stimulation Of

Optogenetic techniques for neural inactivation are valuable for linking neural activity to behavior but they have serious limitations in macaques. To achieve powerful and temporally precise neural inactivation, we used an adeno-associated viral (AAV) vector carrying the channelrhodopsin-2 gene under the control of a Dlx5/6 enhancer, which restricts expression to GABAergic neurons. We tested this approach in the primary visual cortex, an area where neural inactivation leads to interpretable behavioral deficits. Optical stimulation modulated spiking activity and reduced visual sensitivity profoundly in the region of space represented by the stimulated neurons. Rebound firing, which can have unwanted effects on neural circuits following inactivation, was not observed, and the efficacy of the optogenetic manipulation on behavior was maintained across >1000 trials. We conclude that this inhibitory cell-type-specific optogenetic approach is a powerful and spatiotemporally precise neural inactivation tool with broad utility for probing the functional contributions of cortical activity in macaques.

scholarly journals Periaqueductal efferents to dopamine and GABA neurons of the VTA

2017 ◽  
Author(s):  
Niels R. Ntamati ◽  
Meaghan Creed ◽  
Christian Lüscher
Keyword(s):  
Functional Analysis ◽  
Ventral Tegmental Area ◽  
Periaqueductal Gray ◽  
Retrograde Tracing ◽  
The Other ◽  
Projection Neurons ◽  
Cell Type ◽  
Gaba Neurons ◽  
Other Hand ◽  
Cell Type Specific

AbstractNeurons in the periaqueductal gray (PAG) modulate threat responses and nociception. Activity in the ventral tegmental area (VTA) on the other hand can cause reinforcement and aversion. While in many situations these behaviors are related, the anatomical substrate of a crosstalk between the PAG and VTA remains poorly understood. Here we describe the anatomical and electrophysiological organization of the VTA-projecting PAG neurons. Using rabies-based, cell type-specific retrograde tracing, we observed that PAG to VTA projection neurons are evenly distributed along the rostro-caudal axis of the PAG, but concentrated in its posterior and ventrolateral segments. Optogenetic projection targeting demonstrated that the PAG-to-VTA pathway is predominantly excitatory and targets similar proportions of Ih-expressing VTA DA and GABA neurons. Taken together, these results set the framework for functional analysis of the interplay between PAG and VTA in the regulation of reward and aversion.

scholarly journals Npas4 Regulates Excitatory-Inhibitory Balance within Neural Circuits through Cell-Type-Specific Gene Programs

Cell ◽  
2014 ◽  
Vol 157 (5) ◽  
pp. 1216-1229 ◽  
Author(s):  
Ivo Spiegel ◽  
Alan R. Mardinly ◽  
Harrison W. Gabel ◽  
Jeremy E. Bazinet ◽  
Cameron H. Couch ◽  
...  
Keyword(s):  
Neural Circuits ◽  
Specific Gene ◽  
Cell Type ◽  
Cell Type Specific

Viral Strategies for Targeting the Central and Peripheral Nervous Systems

2018 ◽  
Vol 41 (1) ◽  
pp. 323-348 ◽  
Author(s):  
Claire N. Bedbrook ◽  
Benjamin E. Deverman ◽  
Viviana Gradinaru
Keyword(s):  
Nervous System ◽  
Regulatory Elements ◽  
Specific Cell ◽  
Cell Type ◽  
Specific Expression ◽  
Recombinant Viruses ◽  
Neuronal Populations ◽  
Neuroscience Research ◽  
Cell Type Specific Expression ◽  
Cell Type Specific

Recombinant viruses allow for targeted transgene expression in specific cell populations throughout the nervous system. The adeno-associated virus (AAV) is among the most commonly used viruses for neuroscience research. Recombinant AAVs (rAAVs) are highly versatile and can package most cargo composed of desired genes within the capsid's ∼5-kb carrying capacity. Numerous regulatory elements and intersectional strategies have been validated in rAAVs to enable cell type–specific expression. rAAVs can be delivered to specific neuronal populations or globally throughout the animal. The AAV capsids have natural cell type or tissue tropism and trafficking that can be modified for increased specificity. Here, we describe recently engineered AAV capsids and associated cargo that have extended the utility of AAVs in targeting molecularly defined neurons throughout the nervous system, which will further facilitate neuronal circuit interrogation and discovery.

scholarly journals Spatially patterned excitatory neuron subtypes and circuits within the claustrum

2021 ◽  
Author(s):  
Sarah R Erwin ◽  
Brianna N Bristow ◽  
Kaitlin E Sullivan ◽  
Brian Marriott ◽  
Lihua Wang ◽  
...  
Keyword(s):  
Neural Circuits ◽  
Spatial Arrangement ◽  
Retrograde Tracing ◽  
Excitatory Neuron ◽  
Cell Type ◽  
Narrow Region ◽  
Single Cell Rna Sequencing ◽  
Cell Type Specific ◽  
Functional Complexity

The claustrum is a functionally and structurally complex brain region, whose very spatial extent remains debated. Histochemical-based approaches typically treat the claustrum as a relatively narrow region that primarily projects to the neocortex, whereas circuit-based approaches suggest a broader region embedding neocortical and other neural circuits. Here, we took a bottom up, cell-type-specific approach to complement and possibly unite these seemingly disparate conclusions. Using single-cell RNA-sequencing, we found that the claustrum is comprised of two excitatory neuron subtypes that are differentiable from the surrounding cortex. Multicolor retrograde tracing in conjunction with 12-channel multiplexed in situ hybridization revealed a core-shell spatial arrangement of these subtypes, as well as differential projection targets. Thus, the claustrum is comprised of excitatory neuron subtypes with distinct molecular and circuit properties, whose spatial patterns reflect the narrower and broader claustral extents debated in previous research. This subtype-specific heterogeneity likely shapes the functional complexity of the claustrum.

scholarly journals Subcellular sequencing of single neurons reveals the dendritic transcriptome of GABAergic interneurons

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Julio D Perez ◽  
Susanne tom Dieck ◽  
Beatriz Alvarez-Castelao ◽  
Georgi Tushev ◽  
Ivy CW Chan ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Large Population ◽  
Cell Types ◽  
Mrna Localization ◽  
Gabaergic Neurons ◽  
Local Translation ◽  
Single Neurons ◽  
Cell Type ◽  
Cell Type Specific

Although mRNAs are localized in the processes of excitatory neurons, it is still unclear whether interneurons also localize a large population of mRNAs. In addition, the variability in the localized mRNA population within and between cell-types is unknown. Here we describe the unbiased transcriptomic characterization of the subcellular compartments of hundreds of single neurons. We separately profiled the dendritic and somatic transcriptomes of individual rat hippocampal neurons and investigated mRNA abundances in the soma and dendrites of single glutamatergic and GABAergic neurons. We found that, like their excitatory counterparts, interneurons contain a rich repertoire of ~4000 mRNAs. We observed more cell type-specific features among somatic transcriptomes than their associated dendritic transcriptomes. Finally, using cell-type specific metabolic labelling of isolated neurites, we demonstrated that the processes of Glutamatergic and, notably, GABAergic neurons were capable of local translation, suggesting mRNA localization and local translation is a general property of neurons.

scholarly journals Complexity and graded regulation of neuronal cell-type–specific alternative splicing revealed by single-cell RNA sequencing

2021 ◽  
Vol 118 (10) ◽  
pp. e2013056118
Author(s):  
Huijuan Feng ◽  
Daniel F. Moakley ◽  
Shuonan Chen ◽  
Melissa G. McKenzie ◽  
Vilas Menon ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Single Cell ◽  
Neuronal Cell ◽  
Cell Types ◽  
Gabaergic Neurons ◽  
Cell Type ◽  
Differential Splicing ◽  
Single Cell Rna Sequencing ◽  
Hierarchical Levels ◽  
Cell Type Specific

The enormous cellular diversity in the mammalian brain, which is highly prototypical and organized in a hierarchical manner, is dictated by cell-type–specific gene-regulatory programs at the molecular level. Although prevalent in the brain, the contribution of alternative splicing (AS) to the molecular diversity across neuronal cell types is just starting to emerge. Here, we systematically investigated AS regulation across over 100 transcriptomically defined neuronal types of the adult mouse cortex using deep single-cell RNA-sequencing data. We found distinct splicing programs between glutamatergic and GABAergic neurons and between subclasses within each neuronal class. These programs consist of overlapping sets of alternative exons showing differential splicing at multiple hierarchical levels. Using an integrative approach, our analysis suggests that RNA-binding proteins (RBPs) Celf1/2, Mbnl2, and Khdrbs3 are preferentially expressed and more active in glutamatergic neurons, while Elavl2 and Qk are preferentially expressed and more active in GABAergic neurons. Importantly, these and additional RBPs also contribute to differential splicing between neuronal subclasses at multiple hierarchical levels, and some RBPs contribute to splicing dynamics that do not conform to the hierarchical structure defined by the transcriptional profiles. Thus, our results suggest graded regulation of AS across neuronal cell types, which may provide a molecular mechanism to specify neuronal identity and function that are orthogonal to established classifications based on transcriptional regulation.

scholarly journals An extended toolkit for production and use of RVdG-CVS-N2c rabies viral vectors uncovers hidden hippocampal connections

2021 ◽  
Author(s):  
Anton Sumser ◽  
Maximilian Joesch ◽  
Peter Jonas ◽  
Yoav Ben-Simon
Keyword(s):  
Viral Vectors ◽  
Molecular Tools ◽  
Cell Type ◽  
Large Collection ◽  
Neuronal Connectivity ◽  
Retrograde Labeling ◽  
Neuronal Populations ◽  
Wide Range ◽  
Cell Type Specific ◽  
Cortical Microcircuit

From the large collection of molecular tools used to investigate neuronal connectivity, envA-pseudotyped rabies viral vectors (RVdGenvA) uniquely enable cell-type specific, trans-synaptic retrograde labeling. However, widespread use of the powerful and flexible method is to date hindered by low-yield and cumbersome production pipelines. Here, we report the development of new cell lines, which significantly reduce production time while increasing viral titer and eliminating background contamination from native-coat particles. We further show that RVdGenvA-CVS-N2c vectors produced using this system retain their enhanced retrograde-trafficking when compared with SAD-B19 vectors, allowing us to uncover undescribed cortico-hippocampal connections and to monitor activity in a cortical microcircuit of behaving animals. Along with new suites of AAV and RVdG-CVS-N2c vectors, developed to enable retrograde labeling from a wide range of neuronal populations and tailored for diverse experimental requirements, we present here an optimal system for mapping, manipulating and imaging of neuronal circuits.

scholarly journals Acoustically Targeted Chemogenetics for Noninvasive Control of Neural Circuits

2018 ◽  
Author(s):  
Jerzy O. Szablowski ◽  
Brian Lue ◽  
Audrey Lee-Gosselin ◽  
Dina Malounda ◽  
Mikhail G. Shapiro
Keyword(s):  
Neural Circuits ◽  
Brain Regions ◽  
Cell Type ◽  
Spatial Targeting ◽  
Temporal Specificity ◽  
Cell Type Specific ◽  
Orally Bioavailable ◽  
G Protein Coupled ◽  
The Brain ◽  
Spatial Cell

ABSTRACTNeurological and psychiatric diseases often involve the dysfunction of specific neural circuits in particular regions of the brain. Existing treatments, including drugs and implantable brain stimulators, aim to modulate the activity of these circuits, but are typically not cell type-specific, lack spatial targeting or require invasive procedures. Here, we introduce an approach to modulating neural circuits noninvasively with spatial, cell-type and temporal specificity. This approach, called acoustically targeted chemogenetics, or ATAC, uses transient ultrasonic opening of the blood brain barrier to transduce neurons at specific locations in the brain with virally-encoded engineered G-protein-coupled receptors, which subsequently respond to systemically administered bio-inert compounds to activate or inhibit the activity of these neurons. We demonstrate this concept in mice by using ATAC to noninvasively modify and subsequently activate or inhibit excitatory neurons within the hippocampus, showing that this enables pharmacological control of memory formation. This technology allows a brief, noninvasive procedure to make one or more specific brain regions capable of being selectively modulated using orally bioavailable compounds, thereby overcoming some of the key limitations of conventional brain therapies.

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