Climbing Fiber-Mediated Spillover Transmission to Interneurons Is Regulated by EAAT4.
Neurotransmitter spillover is a form of communication not readily predicted by anatomic structure. In the cerebellum, glutamate spillover from climbing fibers recruits molecular layer interneurons in the absence of conventional synaptic connections. Spillover-mediated signaling is typically limited by transporters that bind and reuptake glutamate. Here, we show that patterned expression of the excitatory amino acid transporter 4 (EAAT4) in Purkinje cells regulates glutamate spillover to molecular layer interneurons. Using male and female Aldolase C-Venus knock-in mice to visualize zebrin microzones, we find larger climbing fiber-evoked spillover EPSCs in regions with low levels of EAAT4 compared with regions with high EAAT4. This difference is not explained by presynaptic glutamate release properties or postsynaptic receptor density but rather by differences in the glutamate concentration reaching receptors on interneurons. Inhibiting glutamate transport normalizes the differences between microzones, suggesting that heterogeneity in EAAT4 expression is a primary determinant of differential spillover. These results show that neuronal glutamate transporters limit extrasynaptic transmission in a non-cell-autonomous manner and provide new insight into the functional specialization of cerebellar microzones.SIGNIFICANCE STATEMENT Excitatory amino acid transporters (EAATs) help maintain the fidelity and independence of point-to-point synaptic transmission. Whereas glial transporters are critical to maintain low ambient levels of extracellular glutamate to prevent excitotoxicity, neuronal transporters have more subtle roles in shaping excitatory synaptic transmission. Here we show that the patterned expression of neuronal EAAT4 in cerebellar microzones controls glutamate spillover from cerebellar climbing fibers to nearby interneurons. These results contribute to fundamental understanding of neuronal transporter functions and specialization of cerebellar microzones.
['Malhotra|Shreya|S|', 'Banumurthy|Gokulakrishna|G|', 'Pennock|Reagan L|RL|', 'Vaden|Jada H|JH|', 'Sugihara|Izumi|I|0000-0002-4598-9882', 'Overstreet-Wadiche|Linda|L|', 'Wadiche|Jacques I|JI|']
D000818:Animals D002531:Cerebellum D050581:Excitatory Amino Acid Transporter 4 D019706:Excitatory Postsynaptic Potentials D018698:Glutamic Acid D007395:Interneurons D051379:Mice D011689:Purkinje Cells D013569:Synapses D009435:Synaptic Transmission
spillover cerebellum glutamate EAAT synaptic transmission