NETO1 Guides Development of Glutamatergic Connectivity in the Hippocampus by Regulating Axonal Kainate Receptors

Abstract

Kainate-type glutamate receptors (KARs) are highly expressed in the developing brain where they are tonically activated to modulate synaptic transmission, network excitability and synaptogenesis. NETO proteins are auxiliary subunits that regulate biophysical properties of KARs, however their functions in the immature brain are not known. Here we show that NETO1 guides the development of the rodent hippocampal CA3-CA1 circuitry via regulating axonal KARs. NETO deficiency reduced axonal targeting of most KAR subunits in hippocampal neurons in a subtype independent manner. As an interesting exception, axonal delivery of GluK1c was strongly and selectively impaired in the Neto1 ^-/-, but not Neto2 ^-/- neurons. Correspondingly, the presynaptic GluK1 KAR activity that tonically inhibits glutamate release at immature CA3-CA1 synapses was completely lost in the absence of NETO1 but not NETO2. The deficit in axonal KARs at Neto1 ^-/- neurons resulted in impaired synaptogenesis and perturbed synchronization of CA3 and CA1 neuronal populations during development in vitro. Both these Neto1 ^-/- phenotypes were fully rescued by overexpression of GluK1c, emphasizing the role of NETO1/KAR complex in development of efferent connectivity. Together, our data uncover a novel role for NETO1 in regulation of axonal KARs and identify its physiological significance in development of the CA3-CA1 circuit.

Significance Statement Kainate-type glutamate receptors (KARs) are highly expressed in the developing brain where they modulate synaptic transmission and synaptogenesis. NETO proteins act as auxiliary subunits for KARs, however their functions at the immature circuit are not known. Using a combination of cell biology and electrophysiology in Neto1 and Neto₂ deficient mouse models, we show that NETO proteins regulate axonal delivery of KARs. The deficit in axonal KAR at immature Neto1