Spontaneous exocytosis of single synaptic vesicles generates miniature synaptic currents, which provide a window into the dynamic control of synaptic transmission. To resolve the impact of different factors on the dynamics and variability of synaptic transmission, we recorded miniature excitatory postsynaptic currents (mEPSCs) from cocultures of mouse hippocampal neurons with HEK cells expressing the postsynaptic proteins GluA2, neuroligin 1, PSD-95, and stargazin. Synapses between neurons and these heterologous cells have a molecularly defined postsynaptic apparatus, while the compact morphology of HEK cells eliminates the distorting effect of dendritic filtering. HEK cells in coculture produced mEPSCs with a higher frequency, larger amplitude, and more rapid rise and decay than neurons from the same culture. However, mEPSC area indicated that nerve terminals in synapses with both neurons and HEK cells release similar populations of vesicles. Modulation by the glutamate receptor ligand aniracetam revealed receptor contributions to mEPSC shape. Dendritic cable effects account for the slower mEPSC rise in neurons, whereas the slower decay also depends on other factors. Lastly, expression of synaptobrevin transmembrane domain mutants in neurons slowed the rise of HEK cell mEPSCs, thus revealing the impact of synaptic fusion pores. In summary, we show that cocultures of neurons with heterologous cells provide a geometrically simplified and molecularly defined system to investigate the time course of synaptic transmission and to resolve the contribution of vesicles, fusion pores, dendrites, and receptors to this process.
Recordings from neuron–HEK cell cocultures reveal the determinants of miniature excitatory postsynaptic currents
J. Wan’s and B.A. Weaver’s present address is Departments of Cell and Regenerative Biology and Oncology, Carbone Cancer Center, University of Wisconsin, Madison, WI.
C.-W. Chiang’s present address is Department of Neuroscience, Baylor College of Medicine, Houston, TX.
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Chung-Wei Chiang, Wen-Chi Shu, Jun Wan, Beth A. Weaver, Meyer B. Jackson; Recordings from neuron–HEK cell cocultures reveal the determinants of miniature excitatory postsynaptic currents. J Gen Physiol 3 May 2021; 153 (5): e202012849. doi: https://doi.org/10.1085/jgp.202012849
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