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- Title
CaMKIIα-Positive Interneurons Identified via a microRNA-Based Viral Gene Targeting Strategy.
- Authors
Keaveney, Marianna K.; Rahsepar, Bahar; Hua-an Tseng; Fernandez, Fernando R.; Mount, Rebecca A.; Ta, Tina; White, John A.; Berg, Jim; Xue Han
- Abstract
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 CaMKIIa 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.
- Subjects
VIRAL genes; GENE targeting; GENETIC vectors; GENE expression profiling; INTERNEURONS; INHIBITORY postsynaptic potential; CALBINDIN; COCHLEAR nucleus
- Publication
Journal of Neuroscience, 2020, Vol 40, Issue 50, p9576
- ISSN
0270-6474
- Publication type
Article
- DOI
10.1523/JNEUROSCI.2570-19.2020