Neddylation regulate the development of glutamatergic neurons

The development and function of synapses is orchestrated by a plethora of regulatory mechanisms hierarchically controlling gene expression to protein function. Expanding the proteome, protein post-translational modification are additional key events in neu-rons. In such context, the modification of protein with ubiquitin-like modifier (Ubls) such as the SUMOs, but also other Ubls like NEDD8, have been revealed as major steps in regulating neuronal development and synaptic transmission, either by directly acting lo-cally at synapse or via nuclear-mediated mechanism and the regulation of gene expres-sion. In the present work, we focus on NEDD8 and characterized neuronal morphology, synap-tic transmission and the transcriptional landscape of NEDD8 deficient hippocampal au-taptic neurons. While we observed no major alteration in neuronal morphology, NEDD8-deficient neurons have a decreased pool of ready-releasable synaptic vesicles. This de-fect was likely caused by an imbalance in the expression of vGLUT1/vGLUT2 and de-creased endophilin1 level. Indeed, transcriptome analysis revealed profound defects in the expression of key transcriptional regulators determining glutamatergic neurodevel-opment, indicating that NEDD8 foster the expression of glutamatergic cell fate. We generated a nedd8 conditional knock-out. We cultured primary hippocampal neurons and infected these neurons with either an RFP-expressing virus (as control) or a CRE-expressing virus to ablate NEDD8 expression. After 12 days in culture, RNA were extracted from these culture and their transcriptome was analyzed.