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cAMP Signaling–Mediated Phosphorylation of Diacylglycerol Lipase α Regulates Interaction With Ankyrin-G and Dendritic Spine Morphology

  • Sehyoun Yoon
    Affiliations
    Department of Physiology, Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Kristoffer Myczek
    Affiliations
    Department of Physiology, Northwestern University Feinberg School of Medicine, Chicago, Illinois
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  • Peter Penzes
    Correspondence
    Address correspondence to Peter Penzes, Ph.D.
    Affiliations
    Department of Physiology, Northwestern University Feinberg School of Medicine, Chicago, Illinois

    Department of Psychiatry and Behavioral Sciences, and Pharmacology, Northwestern University Feinberg School of Medicine, Chicago, Illinois

    Center for Autism and Neurodevelopment, Northwestern University Feinberg School of Medicine, Chicago, Illinois
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      Abstract

      Background

      Diacylglycerol lipase α (DAGLα), a major biosynthetic enzyme for endogenous cannabinoid signaling, has emerged as a risk gene in multiple psychiatric disorders. However, its role in the regulation of dendritic spine plasticity is unclear.

      Methods

      DAGLα wild-type or point mutants were overexpressed in primary cortical neurons or human embryonic kidney 293T cells. The effects of mutated variants on interaction, dendritic spine morphology, and dynamics were examined by proximity ligation assay or fluorescence recovery after photobleaching. Behavioral tests and immunohistochemistry were performed with ankyrin-G conditional knockout and wild-type male mice.

      Results

      DAGLα modulated dendritic spine size and density, but the effects of changes in its protein level versus enzymatic activity were different, implicating either a 2-arachidonoylglycerol (2-AG)–dependent or –independent mechanism. The 2-AG–independent effects were mediated by the interaction of DAGLα with ankyrin-G, a multifunctional scaffold protein implicated in psychiatric disorders. Using superresolution microscopy, we observed that they colocalized in distinct nanodomains, which correlated with spine size. In situ proximity ligation assay combined with structured illumination microscopy revealed that DAGLα phosphorylation upon forskolin treatment enhanced the interaction with ankyrin-G in spines, leading to increased spine size and decreased DAGLα surface diffusion. Ankyrin-G conditional knockout mice showed significantly decreased DAGLα-positive neurons in the forebrain. In mice, ankyrin-G was required for forskolin-dependent reversal of depression-related behavior.

      Conclusions

      Taken together, ANK3 and DAGLA, both neuropsychiatric disorder genes, interact in a complex to regulate spine morphology. These data reveal novel synaptic signaling mechanisms and potential therapeutic avenues.

      Keywords

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