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A viral strategy for targeting and manipulating interneurons across vertebrate species.

ABSTRACT: A fundamental impediment to understanding the brain is the availability of inexpensive and robust methods for targeting and manipulating specific neuronal populations. The need to overcome this barrier is pressing because there are considerable anatomical, physiological, cognitive and behavioral differences between mice and higher mammalian species in which it is difficult to specifically target and manipulate genetically defined functional cell types. In particular, it is unclear the degree to which insights from mouse models can shed light on the neural mechanisms that mediate cognitive functions in higher species, including humans. Here we describe a novel recombinant adeno-associated virus that restricts gene expression to GABAergic interneurons within the telencephalon. We demonstrate that the viral expression is specific and robust, allowing for morphological visualization, activity monitoring and functional manipulation of interneurons in both mice and non-genetically tractable species, thus opening the possibility to study GABAergic function in virtually any vertebrate species.

SUBMITTER: Dimidschstein J

PROVIDER: S-EPMC5348112 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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A viral strategy for targeting and manipulating interneurons across vertebrate species.

Dimidschstein Jordane J Chen Qian Q Tremblay Robin R Rogers Stephanie L SL Saldi Giuseppe-Antonio GA Guo Lihua L Xu Qing Q Liu Runpeng R Lu Congyi C Chu Jianhua J Grimley Joshua S JS Krostag Anne-Rachel AR Kaykas Ajamete A Avery Michael C MC Rashid Mohammad S MS Baek Myungin M Jacob Amanda L AL Smith Gordon B GB Wilson Daniel E DE Kosche Georg G Kruglikov Illya I Rusielewicz Tomasz T Kotak Vibhakar C VC Mowery Todd M TM Anderson Stewart A SA Callaway Edward M EM Dasen Jeremy S JS Fitzpatrick David D Fossati Valentina V Long Michael A MA Noggle Scott S Reynolds John H JH Sanes Dan H DH Rudy Bernardo B Feng Guoping G Fishell Gord G

Nature neuroscience 20161031 12

A fundamental impediment to understanding the brain is the availability of inexpensive and robust methods for targeting and manipulating specific neuronal populations. The need to overcome this barrier is pressing because there are considerable anatomical, physiological, cognitive and behavioral differences between mice and higher mammalian species in which it is difficult to specifically target and manipulate genetically defined functional cell types. In particular, it is unclear the degree to ...[more]

PMID: 27798629

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Project description:Single-cell analysis is revealing increasing diversity in gene expression profiles among brain cells. Traditional promotor-based viral gene expression techniques, however, cannot capture the growing variety among single cells. We demonstrate a novel viral gene expression strategy to target cells with specific miRNA expression using miRNA-guided neuron tags (mAGNET). We designed mAGNET viral vectors containing a CaMKIIα promoter and microRNA-128 (miR-128) binding sites, and labeled CaMKIIα+ cells with naturally low expression of miR-128 (Lm128C cells) in male and female mice. Although CaMKIIα has traditionally been considered as an excitatory neuron marker, our single-cell sequencing results reveal that Lm128C cells are CaMKIIα+ inhibitory neurons of parvalbumin or somatostatin subtypes. Further evaluation of the physiological properties of Lm128C cell in brain slices showed that Lm128C cells exhibit elevated membrane excitability, with biophysical properties closely resembling those of fast-spiking interneurons, consistent with previous transcriptomic findings of miR-128 in regulating gene networks that govern membrane excitability. To further demonstrate the utility of this new viral expression strategy, we expressed GCaMP6f in Lm128C cells in the superficial layers of the motor cortex and performed in vivo calcium imaging in mice during locomotion. We found that Lm128C cells exhibit elevated calcium event rates and greater intrapopulation correlation than the overall CaMKIIα+ cells during movement. In summary, the miRNA-based viral gene targeting strategy described here allows us to label a sparse population of CaMKIIα+ interneurons for functional studies, providing new capabilities to investigate the relationship between gene expression and physiological properties in the brain.SIGNIFICANCE STATEMENT We report the discovery of a class of CaMKIIα+ cortical interneurons, labeled via a novel miRNA-based viral gene targeting strategy, combinatorial to traditional promoter-based strategies. The fact that we found a small, yet distinct, population of cortical inhibitory neurons that express CaMKIIα demonstrates that CaMKIIα is not as specific for excitatory neurons as commonly believed. As single-cell sequencing tools are providing increasing insights into the gene expression diversity of neurons, including miRNA profile data, we expect that the miRNA-based gene targeting strategy presented here can help delineate many neuron populations whose physiological properties can be readily related to the miRNA gene regulatory networks.

Dataset Information

A viral strategy for targeting and manipulating interneurons across vertebrate species.

Publications

A viral strategy for targeting and manipulating interneurons across vertebrate species.

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