Journal: eLife
Article Title: Alcohol attenuates CRF-induced excitatory effects from the extended amygdala to dorsostriatal cholinergic interneurons
doi: 10.7554/eLife.107145
Figure Lengend Snippet: ( A ) Sample images of GFP-labeled CINs in the striatum of a ChAT-eGFP mouse. Scale bar: 0.5 mm, 50 μm (inset). ( B ) Bath application of CRF (100 nM) increased the spontaneous firing of dorsal striatal CINs in cell-attached electrophysiological recordings. This effect was prevented by pretreatment with the CRFR1 antagonist NBI 35695 (5 μM). n=7 cells from 3 mice (7/3) for CRF and 6/3 for CRF plus antagonist recordings. ( C ) Data showing that CRF significantly increases the firing from baseline, and the firing frequency in the presence of CRF following pretreatment with CRFR1 antagonist is significantly lower. ***p<0.001 by paired t-test, ***p<0.001 by Mann-Whitney test. n=7 cells from 3 mice. ( D ) Sample images of ACh sensor fluorescence in dorsal striatal slices before and during bath application of CRF (100 nM). AAV-hSyn-GRAB ACh4m was infused into the dorsal striatum of wild-type mice, and live-tissue confocal imaging was conducted 2 weeks post-infusion. Scale bar: 10 μm for left and right. ( E ) Sample trace of spontaneous ACh release events (indicated by red arrows, top). Bath application of CRF increased ACh sensor fluorescence (bottom). ( F ) Summary data showing a significant increase in ACh sensor fluorescence following CRF application. *p<0.05 by Mann-Whitney test. n=7 slices from 7 mice. ( G ) Schematic of cell-attached electrophysiological recordings from dorsal striatal CINs in CRF-Cre;Ai32;ChAT-eGFP mice, with simultaneous optogenetic stimulation of CRF + fibers using blue light (470 nm, 2 ms, 50 Hz, 60 s). ( H ) Sample trace showing the increase in CIN firing frequency during blue light stimulation of CRF + fibers. ( I ) Data demonstrating a significant and reversible increase in CIN firing frequency during optogenetic stimulation, *p<0.05, ***p<0.001 by one-way RM ANOVA. n=13 neurons from 8 animals. ( J ) Sample trace showing no change in CIN firing frequency during blue light stimulation of CRF + fibers when CRFR1 antagonist (antalarmin hydrochloride, 2 μM) was bath applied. (K) Data demonstrating no significant change in CIN firing frequency during optogenetic stimulation. n=9 neurons from 3 mice. Data are presented as mean ± SEM.
Article Snippet: ChAT-eGFP (stock 007902), ChAT-Cre (stock 031661), Drd1a-tdTomato (D1-tdT, stock 016204), Ai32 (stock 012569), CRH-ires-CRE (stock 012704), and C57BL/6J (stock 000664) mice were purchased from The Jackson Laboratory ( ; ; ; ; ).
Techniques: Labeling, MANN-WHITNEY, Fluorescence, Imaging
