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. 2019 May 22;5(5):eaaw5141. doi: 10.1126/sciadv.aaw5141

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Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 5 — (A) A labella-brain preparation. (B) Patch-clamp recordings of GMR58H01 MNs. Left: SEZ neurons labeled by GMR58H01-Gal4 (top) and four pairs of SEZ MNs labeled by an intersection between vGlut-FLP and GMR58H01-Gal4 (bottom). “a,” “b,” “c,” and “d” mark the four pairs of MNs with similar positions as those MNs at the bottom. Middle: Illustration of simultaneous optogenetic activation of peg MSNs and patch-clamp recordings of GMR58H01 MNs. Right: Electrical responses of GMR58H01 MNs under cell-attached and perforated patch-clamp recordings and collective data. (C) GRASP between labellar MSNs and GMR58H01 neurons. Left: GRASP signal between labellar MSNs and GMR58H01 neurons (top); no GRASP signal in the absence of GMR58H01 driver (bottom). Right: GRASP signal in SEZ (expanded from top left panel). (D) Monosynaptic transmission from peg MSNs to GMR58H01 MNs. Left: Illustration of local optogenetic activation of labella and patch-clamp recordings on MNs in the brain (top) and spike firing in peg MSNs by optogenetic activation (bottom). Middle: Optogenetic activation of peg MSNs triggers an inward current in GMR58H01 MNs (top) and at the expanded time scale (bottom). Right: Collective data of synaptic delay from peg MSNs to GMR58H01 MNs. Light stimulation: 2 ms, 617 nm. (E) Bitter GRNs enhance peg MSN–driven proboscis retraction. Top: Proboscis retraction is more sensitive by coactivating peg MSNs and bitter GRNs. Light stimulation: 100 ms, 617 nm. Bottom: The latency of proboscis retraction is reduced by coactivation of peg MSNs and bitter GRNs. Light stimulation: 617 nm, 1.75 mW/cm², 1 s. ***P < 0.001. (F) Inward currents of GMR58H01 MNs triggered by peg MSNs and bitter GRNs. Top: Current of GMR58H01 MNs driven by optogenetic activation of peg MSNs, bitter GRNs, and peg MSNs and bitter GRNs together. Bottom: Collective data. CAF, 20 mM. Light stimulation: 617 nm, 1.75 mW/cm². *P < 0.05 and ***P < 0.001. (G) Activation of GMR58H01 MNs by bitter GRNs independently of peg MSNs. Inward currents of GMR58H01 MNs driven by bitter GRNs (top) is intact when synaptic transmission of peg MSNs is blocked (middle); collective data (bottom). CAF, 20 mM.