The role of agrin, Lrp4 and MuSK during dendritic arborization and synaptogenesis in cultured embryonic CNS neurons

• Published in: Developmental biology 2019-01, Vol.445 (1), p.54-67
• Main Authors: Handara, Gerry, Hetsch, Florian J.A., Jüttner, René, Schick, Anna, Haupt, Corinna, Rathjen, Fritz G., Kröger, Stephan
• Format: Article
• Language: English
• Subjects: Autapses; Dendritic arborization; GABAA receptor; Gephyrin; PSD-95; Synaptogenesis
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1016/j.ydbio.2018.10.017

The role of agrin, Lrp4 and MuSK, key organizers of neuromuscular synaptogenesis, in the developing CNS is only poorly understood. We investigated the role of these proteins in cultured mouse embryonic cortical neurons from wildtype and from Lrp4- and MuSK-deficient mice. Neurons from Lrp4-deficient mice had fewer but longer primary dendrites and a decreased density of puncta containing excitatory and inhibitory synapse-associated proteins. Neurons from MuSK-deficient mice had an altered dendritic branching pattern but no change in the density of puncta stained by antibodies against synapse-associated proteins. Transfection of TM-agrin compensated the dendritic branching deficits in Lrp4-deficient but not in MuSK-deficient neurons. TM-agrin transfection increased the density of excitatory synaptic puncta in MuSK-deficient but not in Lrp4-deficient mice and reduced the number of inhibitory synaptic puncta irrespective of MuSK and Lrp4 expression. Addition of purified soluble agrin to microisland cultures of cortical neurons revealed an Lrp4-dependent increase in the size and density of glutamatergic synaptic puncta and in mEPSC but not in mIPSC frequency and amplitude. Thus, agrin induced an Lrp4-independent increase in dendritic branch complexity, an Lrp4-dependent increase of excitatory synaptic puncta and an Lrp4- and MuSK-independent decrease in the density of puncta containing inhibitory synapse-associated proteins. These results establish selective roles for agrin, Lrp4 and MuSK during dendritogenesis and synaptogenesis in cultured CNS neurons. •Lrp4- and MuSK-deficient embryonic neurons have a reduced dendritic complexity.•TM-agrin transfection restored complexity in Lrp4- but not in MuSK-deficient neurons.•TM-agrin overexpression caused Lrp4-dependent increase in excitatory synapses.•TM-agrin overexpression caused Lrp4-dependent decrease in inhibitory synapses.•Soluble agrin-mediated increase in mEPSC frequency and amplitude depended on Lrp4.