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Functional coupling of Tmem74 and HCN1 channels regulates anxiety-like behavior in BLA neurons

Abstract

Anxiety disorders are the most prevalent psychiatric disorders, but their pathogenic mechanism remains poorly understood. Here, we report that transmembrane protein 74 (TMEM74), which contains two putative transmembrane domains and exhibits high levels of mRNA in the brain, is closely associated with the pathogenesis of anxiety disorders. TMEM74 was decreased in the serum of patients with anxiety and the basolateral amygdaloid nucleus (BLA) in chronic stress mice. Furthermore, genetic deletion of Tmem74 or selective knockdown of Tmem74 in BLA pyramidal neurons resulted in anxiety-like behaviors in mice. Whole-cell recordings in BLA pyramidal neurons revealed lower hyperpolarization-activated cation current (Ih) and greater input resistance and excitability in Tmem74−/− neurons than in wild-type neurons. Accordingly, surface expression of hyperpolarization-activated cyclic nucleotide-gated 1 (HCN1) channels was also lower in the BLA of Tmem74−/− mice. The Ih current blocker ZD7288 mimicked these effects in BLA pyramidal neurons in wild-type mice but not in Tmem74−/− mice. Consistent with the improvement in anxiety-like behaviors, Tmem74 overexpression restored HCN1 channel trafficking and pyramidal neuron excitability in the BLA of Tmem74−/− and chronic stress mice. Mechanistically, we demonstrate that interactions between Tmem74 and HCN1 are physiologically relevant and that transmembrane domain 1 (TM1) is essential for the cellular membrane localization of Tmem74 to enhance Ih. Together, our findings suggest that Tmem74 coupling with HCN1 acts as a critical component in the pathophysiology of anxiety and is a potential target for new treatments of anxiety disorders.

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pCLAMP 10.3 software (Molecular Devices, Sunnyvale, CA, USA) and Matlab software (MathWorks, Natick, MA, USA) were used for electrophysiology data analysis. All codes are available from the authors upon reasonable request.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2016YFE0125400 to F.H.); The State Key Program of National Natural Science of China (grant 81730101 to F.H.); National Natural Science Foundations of China (81573411 to F.H., 81673415 to YM.L., 81601177 to QZ.C.); Science and Technology Commission Foundation of Hangzhou (20172016A05 to YM.L.). We thank the Human Protein Atlas for support with RNA and protein expression data.

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Shao, LX., Jiang, Q., Liu, XX. et al. Functional coupling of Tmem74 and HCN1 channels regulates anxiety-like behavior in BLA neurons. Mol Psychiatry 24, 1461–1477 (2019). https://doi.org/10.1038/s41380-019-0402-8

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