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. 2022 Feb 22;13:998. doi: 10.1038/s41467-022-28493-4

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© The Author(s) 2022

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. 1 — a The experimental strategy of recording the activity of extratelencephalic projection neurons in the mPFC with fiber photometry. b An example image showed the virus injection site and the position of the optical fiber. c The experimental procedure of novel object recognition test. d An example showed the trace of the calcium signal and the time point of the object interaction during object recognition test. e Heatmap of Ca²⁺ transients of extratelencephalic projection neurons in PL of Fezf2-CreER mice during object interactions. f Average plots across different trials of individual animals show that object interactions activated extratelencephalic projection neurons in mPFC. g Heatmap of Ca²⁺ transients of extra telencephalic projection neurons in PL of AD _Fezf2-CreER mice during object interactions. h Average plots across different trials of individual animals show that object interactions activated extratelencephalic projection neurons in mPFC of 5×FAD mice. i Heatmap of Ca²⁺ transients of extratelencephalic projection neurons in PL of Fezf2-CreER mice during object interactions. The GFP was expressed in the PL instead of GCaMP6s. j Average plots across different trials of individual animals show that object interactions did not evoke clear response in GFP positive neurons. k The average plots of the calcium response from different animals. l. The quantification of area under curve (AUC) in k. two-tailed unpaired t test was used, Fezf2-CreER mice, n = 7 animals; AD_Fezf2-CreER, n = 5 animals. m Immunostaining against c-fos to validate the extratelencephalic projection neurons were activated in the object recognition test. White arrow indicates c-fos, yellow arrow indicates Aβ plaques. n Quantification of the number of extratelencephalic projection neurons that expressed GCaMP in the mPFC. n = 4 animals. One-way repeated-measures (RM) ANOVA with Tukey’s post hoc test. o Quantification of number of c-fos positive extratelencephalic projection neurons in the mPFC. one-way repeated-measures (RM) ANOVA with Tukey’s post hoc test, n = 4 animals. p Quantification of the percentage of c-fos positive extratelencephalic projection neurons in the mPFC. one-way repeated-measures (RM) ANOVA with Tukey’s post hoc test, n = 4 animals. Scale bar in b is 500 μm. Scale bar in m is 100 μm. All data are listed as the Mean ± SEM. The color bars in the figure indicated df/f (%). NOR, novel object recognition test. TM, tamoxifen.