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. 2022 Jan 17;13:339. doi: 10.1038/s41467-022-27977-7

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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. 2 — a Schematic of fluorescent bead (retrobeads) injection into PFC and retrograde labelling in transverse hippocampal slices prepared for recordings. b Example reconstructions of superficial (blue) and deep (dark grey) PFC-projecting hippocampal neurons. Scale bar = 100 µm, dotted line represents alveus. Images representative of 7 superficial and 6 deep neurons. c Sholl analysis showing average dendritic length of apical (top) and basal (bottom) dendrites with increasing distance from the alveus (lateral) for superficial (blue) and deep (grey) layer vH-PFC neurons. Note that each population samples a distinct range of the radial hippocampal axis. (Apical: repeated measures ANOVA, Main effect of group: F_(35,385) = 1.549, p = 0.027; basal: repeated measures ANOVA with Greenhouse-Geisser correction, Main effect of group: F_(33,367) = 1.701, p = 0.179; n = 7 (superficial), n = 6 (deep) neurons from 6 mice). d Left, quantification of the distance from the farthest dendrite tip to the soma (two-sided Mann Whitney, U = 35.5, *p = 0.045). Right, quantification of the distance of the apical bifurcation from the soma in superficial (Sup) and Deep neurons (two-sided Mann Whitney, U = 38.5, *p = 0.015, n = 7 (superficial), n = 6 (deep) neurons from 6 mice). e Left, example response of a superficial layer PFC-projecting cell in response to a current injection of 140 pA. Top right, detail of first 50 ms of current injection. Bottom right, response to a current injection of –160 pA. Scale bars = 100 ms, 20 mV; 10 ms, 20 mV; 100 ms, 20 mV. f As in (e) but for a neighboring deep layer PFC-projecting neuron. Note burst firing in response to current injection, and lower level of voltage sag after negative current injection. g Quantification of number of action potentials elicited by somatic current injection in superficial (blue) and deep (black) neurons (Repeated measures ANOVA with Greenhouse-Geisser correction, Interaction between group and current step: F_{(1.4, 44.5)} = 4.3, *p = 0.032, n = 22 (sup) and 26 (deep) from 12 mice). h Left, quantification of the proportion of bursting neurons after positive current injections (Fishers Exact test, Odds ratio = 12.5, *p = 0.009). Right, quantification of sag amplitude after negative (–160 pA) current injections (two-sided Mann Whitney, U = 39.5, *p = 0.002, n = 22 (sup) and 26 (deep) from 12 mice). Box plots show median, 75th (box) and 95th (whiskers) percentile. Line plots show mean (line) and SEM (shading and error bars).