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Image Search Results
Journal: eLife
Article Title: Cestode larvae excite host neuronal circuits via glutamatergic signalling
doi: 10.7554/elife.88174.3
Figure Lengend Snippet: Figure 4. Taenia crassiceps products contain excitatory amino acids detected using iGluSnFR. (A) Schematic of the experimental setup for glutamate detection by the genetically encoded fluorescent glutamate reporter iGluSnFR, including puff pipette and sCMOS camera for imaging following excitation using a 475/28 nm LED-based light engine. (B) Left, iGluSnFR-expressing neuron with the region of interest (red dashed line) used to calculate dF/F trace (right) during a 20-ms T. crassiceps homogenate (T. crass. hom.) puff (orange arrow). (C) As in ‘B’ but with a 20-ms T. crassiceps excretory/ secretory (T. crass. E/S) product puff (orange arrow). (D) Schematic of experimental setup as in ‘A’ but with glutamate or aspartate application via puff pipette. (E) iGluSnFR fluorescence just after a 20-ms puff of artificial cerebrospinal fluid (aCSF) containing 100 μM glutamate (blue arrow). (F) As in ‘E’ but with iGluSnFR fluorescence during a 20-ms puff of aCSF containing 100 μM aspartate (purple arrow). (G) Population data comparing peak dF/F shifts after a 20-ms puff of aCSF (as a negative control), T. crass. hom., T. crass. E/S products, aCSF containing 100 μM glutamate (as a positive control), or aCSF containing 100 μM aspartate (as a positive control), values with medians ± IQR, *p ≤ 0.05, ****p ≤ 0.0001. (H) Schematic of whole-cell patch-clamp recording in current-clamp mode from a CA3 pyramidal neuron using a cesium-based internal and in the presence of 2 μM tetrodotoxin (TTX). 20-ms puff of aCSF containing 100 μM glutamate produces a significant depolarizing shift in membrane potential (top trace), a similar puff but with 100 μM aspartate elicited a similar response (bottom trace). (I) Population data comparing peak membrane potential shift after a 20-ms puff of T. crass. hom., T. crass. E/S, aCSF containing 100 μM glutamate, and aCSF containing 100 μM aspartate. Values with medians ± IQR; NS = not significant.
Article Snippet: For glutamate imaging, excitation was generated using an
Techniques: Transferring, Imaging, Expressing, Fluorescence, Negative Control, Positive Control, Patch Clamp, Membrane