Journal: Molecular psychiatry
Article Title: Inhibitory regulation of calcium transients in prefrontal dendritic spines is compromised by a nonsense Shank3 mutation
doi: 10.1038/s41380-020-0708-6
Figure Lengend Snippet: (a) Coronal histological section, showing the extent of AAV-mediated expression of GCaMP6f in the mouse medial prefrontal cortex. Cg1, cingulate cortex. M2, secondary motor cortex. PrL, prelimbic cortex. (b) Left and middle, schematic of experimental setup and location. Right, an in vivo two-photon image of GCaMP6f-expressing apical dendritic spines in layer 1 of Cg1/M2 regions of medial prefrontal cortex. Two dendritic spines, s1 and s2, were identified based on their morphology as protrusions attached to the dendrite. Arrows, spines s1 and s2. Dashed lines, the dendritic shaft adjacent to the spines. (c) A scatter plot of the fluorescence transients (ΔF/F) measured from spine s1 against the ΔF/F measured from the adjacent dendritic shaft. Each open circle represents an image frame. Line, a least-squares regression line forced through the origin. (d) Top, example segment of ΔF/F signal recorded from the dendritic shaft (solid green line). Middle, ΔF/F signals recorded from the two dendritic spines, s1 and s2 (solid yellow lines), plotted along with the predicted non-local component of the spine calcium signal – the dendritic shaft calcium signal scaled based on the least-squares regression (broken green lines). Bottom, subtracting the scaled dendritic shaft ΔF/F signal from the dendritic spine ΔF/F signals to get an estimate of the synaptic calcium signals for spines s1 and s2 (solid black lines). (e) In vivo two-photon image of GCaMP6s-expressing long-range axons from retrosplenial cortex and jRGECO1a-expressing dendrites in the medial prefrontal cortex. Note a pair of apposing axonal bouton (arrowhead) and dendritic spine (arrow). (f) Fluorescence traces for the bouton and opposing spine (either with subtraction or no subtraction of dendritic shaft contribution) as denoted by the arrow and arrowhead respectively in (a). (g) Cumulative plot of the conditional probability of detecting a calcium event in an axonal bouton given detection of a calcium event in an apposing dendritic spine (data with subtraction: 0.65 ± 0.01, data without subtraction: 0.329 ± 0.004, shuffled: 0.144 ± 0.004, mean ± s.e.m.; P = 8 × 10 −43 , paired t-test). n = 45 bouton-spine pairs from 3 animals.
Article Snippet: For the two-color imaging experiment, two viruses were injected in the same animal: AAV1-Syn-NES-jRGECO1a-WPRE-SV40 (Addgene; 1 × 10 13 GC/mL titer, 294.4 nL) in Cg1/M2, and AAV1-Syn-GCaMP6s-WPRE-SV40 (Penn Vector Core; 3 × 10 13 GC/mL titer, 294.4 nL) in RSC.
Techniques: Expressing, In Vivo, Fluorescence
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