Journal: bioRxiv
Article Title: Corticotropin-releasing hormone signaling from prefrontal cortex to lateral septum supports social novelty preference
doi: 10.1101/2022.03.15.484224
Figure Lengend Snippet: a. DIC image of rdLS during patch-clamp recording. Scale bar: 500 µm. b. Example traces of IPSCs before or 15 min after application of 300 nM stressin-1. c. Frequency of IPSCs. d. Amplitude of IPSCs. e. IPSCs area under the curve. For c-e, points are obtained from individual cells recorded from separate slices in 6 mice. f. C57BL/6J wild-type mice injected in rdLS with AAV2/1 Syn.GCaMP6f and implanted with an optical ferrule above rdLS. Implanted mice were presented with novel and familiar mice. g. Interaction time during social presentation. Dots are from 9 recording sessions using 5 mice. h. Average peak amplitude of the z-score during presentation of a novel or familiar mouse. Paired t test: p = 0.007. i. Frequency of events during presentation of a novel or a familiar mouse. Paired t test: p = 0.6. j. Discrimination index for social familiarity preference calculated from z-scores. One-sample t tests compared to 0: p = 0.006. k. Immunohistochemistry images of c-Fos labelling in rdLS following social presentation with a novel or familiar mouse (same experiment than ). Scale bars: 500 µm. l. Density of rdLS cells positive for c-Fos. We made one observation on each side of a rLS section. 4 mice per group. Unpaired t test, p = 0.0003. m. Density of rdLS cells positive for c-Fos vs. interaction time during social interaction. Each point represents one mouse. n. Percentage of layer 2/3 ILA CRH cells positive for c-Fos (cf. ) vs. density of rdLS cells positive for c-Fos following social interaction. Each point represents a mouse. o-p. Immunohistochemistry images of c-Fos labelling in ILA (o) and rdLS (p). CRH-Cre mice injected in ILA with AAV2/9 CMV-DIO-(mCherry-U6)-shRNA(anti- Crh ) or AAV2/9 CMV-DIO-(mCherry-U6)-shRNA(scrambled) were presented with a familiar mouse for 2 min before being processed for immunohistochemistry. Scale bars: 300 µm. q. Duration of interaction during familiar presentation. Each point is one mouse. Unpaired t test, p = 0.001. r. Percentage of layer 2/3 ILA CRH cells positive for c-Fos in layer 2/3 of ILA. Each point corresponds to each side of 2 sections. 9 mice per group. s. Density of rdLS cells positive for c-Fos. We made one observation on each side of a rLS section. 9 mice per group. Unpaired t test, p < 0.0001. t. Percentage of layer 2/3 ILA CRH cells positive for c-Fos vs. density of rdLS cells positive for c-Fos following social interaction with a familiar mouse. Each point represents one mouse. u. C57BL/6J wild-type mice were injected with AA2/2 hSyn1.hChR2(H134R)-mCherry or AA2/2 hSyn1.mCherry as control and an optical fiber was implanted above the injection site. Mice were then presented to a familiar mouse for 2 min meanwhile 450 nm light was applied (20 Hz, 1 ms). Mice were also run without light as additional controls. Scale bar: 1 mm. v. Total interaction time with familiar mouse. Each point represents one mouse. One-way ANOVA: F 3,32 = 7.05, p = 0.0009. Dunnett’s multiple comparisons tests: ChR-light vs. YFP-no light p = 0.0005, ChR-light vs. ChR-no light p = 0.006, ChR-light vs. YFP-light p = 0.01. w. Average duration of each bout of social interaction. Each point represents one mouse. One-way ANOVA: F 3,31 = 10.62, p < 0.0001. Dunnett’s multiple comparisons tests: ChR-light vs. YFP-no light p = 0.0001, ChR-light vs. ChR-no light p = 0.0001, ChR-light vs. YFP-light p = 0.002. x. Total distance travelled. Each point represents one mouse. y. Schematic of the vCA1-ILA CRH -rdLS circuit. For the entire figure, bar graphs represent mean ± S.E.M.
Article Snippet: After 10 min of stable baseline recording, stressin-1 (300 nM, Tocris # 1608) was applied following a 1:1000 dilution from stock solution into the ACSF.
Techniques: Patch Clamp, Injection, Immunohistochemistry, shRNA
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