Abstract
GABA-A receptors (GABAARs) are crucial for development and function of the brain. Altered GABAergic transmission is hypothesized to be involved in neurodevelopmental disorders. Recently, we identified Shisa7 as a GABAAR auxiliary subunit that modulates GABAAR trafficking and GABAergic transmission. However, the underlying molecular mechanisms remain elusive. Here we generated a knock-in (KI) mouse line that is phospho-deficient at a phosphorylation site in Shisa7 (S405) and combined with electrophysiology, imaging and behavioral assays to illustrate the role of this site in GABAergic transmission and plasticity as well as behaviors. We found that expression of phospho-deficient mutants diminished α2-GABAAR trafficking in heterologous cells. Additionally, α1/α2/α5-GABAAR surface expression and GABAergic inhibition were decreased in hippocampal neurons in KI mice. Moreover, chemically induced inhibitory long-term potentiation was abolished in KI mice. Lastly, KI mice exhibited hyperactivity, increased grooming and impaired sleep homeostasis. Collectively, our study reveals a phosphorylation site critical for Shisa7-dependent GABAARs trafficking which contributes to behavioral endophenotypes displayed in neurodevelopmental disorders.
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Acknowledgements
We are grateful to all members from the Lu laboratory for critical comments on the manuscript. We thank Daniel Abebe at NIH/NICHD for assistance with behavioral tests.
Funding
This work was supported by the NIH/NINDS Intramural Research Program (to WL), the NIH/NEI Intramural Research Program (to LD), Postdoctoral Fellowship from the NIH Center on Compulsive Behaviors (to RDS) and NINDS Diversity Training Fellowship (to DC).
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KW, RDS, and WL designed the project, and WL supervised the project. KW performed imaging and electrophysiological experiments. KW and WH performed biochemical experiments. KW and RDS performed behavioral assays. DC recorded GABA-evoked currents in HEK293T cells and neurons. QT performed neuronal cultures. LD generated Shisa7 S405A mutant mice. KW, RDS, and WL wrote the manuscript, and all authors read and commented on the manuscript.
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Wu, K., Shepard, R.D., Castellano, D. et al. Shisa7 phosphorylation regulates GABAergic transmission and neurodevelopmental behaviors. Neuropsychopharmacol. 47, 2160–2170 (2022). https://doi.org/10.1038/s41386-022-01334-0
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DOI: https://doi.org/10.1038/s41386-022-01334-0
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