Figure 1: Thy1 lines mark BLA population that is active during expression of fear extinction. | Nature Communications

Figure 1: Thy1 lines mark BLA population that is active during expression of fear extinction.

From: Molecular characterization of Thy1 expressing fear-inhibiting neurons within the basolateral amygdala

Figure 1

(a) Thy1-eYFP, (b) Thy1-eNpHR, (c) Thy1-ChR2, each have strong expression enriched in the BLA. (d) Cre recombinase driven by the Thy1.2 cassette in the Thy1-cre mouse, when visualized via infusion of AAV- hSyn-DIO-rM3D(Gs)-mCherry, marks the same regional population as marked by the Thy1-eYFP mouse. (e) Representative images of a field of view analysed for c-fos staining in the BLA of Thy1-YFP mouse after fear expression. Co-localization was manually scored at × 40 magnification, × 10 images are for reference. Upper panels: YFP+ expression in BLA of examined Thy-1 YFP mice. Green neurons in upper panels are YFP+ neurons while black c-fos+ nuclei are depicted in lower panels. Fields for analysis were chosen at random within strongly YFP expressing area. Fields of were visually inspected for co-labelling of c-fos and YFP. Example neurons identified: arrowhead indicates c-fos+/YFP+ cell, while asterisk indicates c-fos+/YFP− cell. (f) Quantification of co-labelling of YFP or non-YFP-marked cells with c-fos in Thy1-eYFP mouse. Counts represented as number of positive events per field analysed. BLA sections were analysed after (1) untrained home cage controls (HC), (2) fear conditioning (FC), (3) fear extinction (FE) or (4) fear extinction retention (FR). Thy1-eYFP+ neurons are c-fos+ significantly more often after fear extinction testing versus home cage, while eYFP-marked neurons stain for c-fos significantly more often during fear expression (two-way ANOVA, F(3,40) P=0.0017, when significant main or interaction effects were found by the ANOVAs, Sidak’s post hoc tests were carried out to locate simple effects. Sidak’s multiple comparisons versus home cage: *P<0.05, *P<0.005, error bars indicate ±s.e.m. Sidak’s multiple comparison post hoc analysis: YFP+ HC versus YFP− FE P<0.05, YFP+ FC versus YFP− FE P<0.05, YFP+ FE versus YFP− FE: P<0.05, YFP− HC versus YFP− FE P<0.05, YFP− FE versus YFP− FR: P<0.05, error bars indicate mean±s.e.m.). (g–i) Infusion of AAV-EF1a-DIO mCherry into double transgenic Thy1-eYFP/Thy1-Cre mouse allows co-localization of neurons marked by each transgene. Arrows indicate neurons with co-localization. Asterisks indicate neuron marked only by mCherry. Scale bar, 100 μm (a–d, e upper left and lower left); scale bar, 20 μm for (g–i, e upper right and lower right).

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