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. 2021 May 4;12:2517. doi: 10.1038/s41467-021-22730-y

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Timeline of the approach-avoidance conflict test during single-unit recordings from photoidentified aPVT^CRF neurons. b Diagram showing the injection of viral mix containing AAV-CRF-Cre and AAV-ChR2-DIO, and the implantation of optrode in aPVT. c–e Photoidentification of aPVT^CRF neurons. c Representative aPVT^CRF neuron responsive to laser illumination (Z-score >3.29, P < 0.001, red dotted line, see details in Methods). d Cells with photoresponse latencies <12 ms were classified as aPVT^CRF neurons (black bars, n = 26 out of 96 recorded neurons), whereas cells with photoresponse latencies >12 ms (white bars, n = 5 neurons) or non-responsive to the laser (n = 65 neurons, not shown) were classified as non-identified aPVT neurons (aPVT^non-ident, n = 70 out of 96 recorded neurons). e Raster plot and firing rate of a representative aPVT^CRF neuron responding to a 5 Hz train of laser stimulation. Inset: Raster plot and firing rate time-locked for laser onset. Vertical blue bars: laser onset. Bins of 1 ms. f Relative frequency histogram showing the baseline firing rate of aPVT^CRF neurons and aPVT^non-identif neurons. g (Top) Schematic of the food-cue-evoked responses. (Bottom) Percentage of aPVT^CRF and aPVT^non-ident neurons that were responsive to food cues (green bars) before (left) and during the conflict (right). aPVT^CRF neurons showed more food-cue responses during the conflict test, when compared to aPVT^non-ident neurons (Fisher’s exact test; aPVT^CRF neurons: 39%, 10 out of 26; aPVT^non-ident neurons: 15%, 11 out of 70 neurons, P = 0.039). h–i Average peristimulus time histograms of all photoidentified aPVT^CRF neurons showing h excitatory or i inhibitory food-cue responses during the conflict (red or blue bars, respectively) or the same neurons before the conflict (gray bars). j (Top) Schematic of the spontaneous activity recordings. (Bottom) Percentage of aPVT^CRF and aPVT^non-ident neurons that changed their baseline spontaneous activity (30 s pre vs. 30 s post) exclusively in food-seeking phase, cat odor phase, conflict phase (30 min), in more than one phase (nonselective), or did not change. No differences were observed between the two groups (Fisher’s exact test, all P’s > 0.05). k (Top) Schematic of the recordings during lever presses-evoked responses. (Bottom) Percentage of aPVT^CRF and aPVT^non-ident neurons that were responsive to lever presses (pink bars) before the conflict phase. l (Top) Schematic of the recordings during dish-entry-evoked responses. (Bottom) Percentage of aPVT^CRF and aPVT^non-ident neurons that were responsive to rewarded dish entries (orange bars) before the conflict phase. No differences were observed between the two groups (Fisher’s exact test, all P’s > 0.05). A total of eight rats were used.