Simultaneous impairment of neuronal and metabolic function of mutated gephyrin in a patient with epileptic encephalopathy

Abstract

© 2015 EMBO.Synaptic inhibition is essential for shaping the dynamics of neuronal networks, and aberrant inhibition plays an important role in neurological disorders. Gephyrin is a central player at inhibitory postsynapses, directly binds and organizes GABAA and glycine receptors (GABAARs and GlyRs), and is thereby indispensable for normal inhibitory neurotransmission. Additionally, gephyrin catalyzes the synthesis of the molybdenum cofactor (MoCo) in peripheral tissue. We identified a de novo missense mutation (G375D) in the gephyrin gene (GPHN) in a patient with epileptic encephalopathy resembling Dravet syndrome. Although stably expressed and correctly folded, gephyrin-G375D was non-synaptically localized in neurons and acted dominant-negatively on the clustering of wild-type gephyrin leading to a marked decrease in GABAAR surface expression and GABAergic signaling. We identified a decreased binding affinity between gephyrin-G375D and the receptors, suggesting that Gly375 is essential for gephyrin-receptor complex formation. Surprisingly, gephyrin-G375D was also unable to synthesize MoCo and activate MoCo-dependent enzymes. Thus, we describe a missense mutation that affects both functions of gephyrin and suggest that the identified defect at GABAergic synapses is the mechanism underlying the patient's severe phenotype. Synopsis: Aberrant inhibitory synaptic transmission is linked to brain disorders including epilepsy, autism and intellectual disability. A de novo mutation was identified in the gephyrin gene in a patient with epileptic encephalopathy. This mutation disrupts gephyrin postsynaptic and molybdenum cofactor biosynthesis functions. Gephyrin-G375D mutation in a patient with Dravet-like syndrome and intellectual disability does not cluster at inhibitory postsynapses and acts dominant-negatively on wild-type gephyrin Postsynaptic clustering and inhibitory signaling is diminished by gephyrin-G375D The mutation weakens the interaction between gephyrin and inhibitory GABAA and glycine receptors Gephyrin-G375D is enzymatically inactive and unable to synthesize the molybdenum cofactor, thus the mutation affects both gephyrin functions Aberrant inhibitory synaptic transmission is linked to brain disorders including epilepsy, autism and intellectual disability. A de novo mutation was identified in the gephyrin gene in a patient with epileptic encephalopathy. This mutation disrupts gephyrin postsynaptic and molybdenum cofactor biosynthesis functions.