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. 2019 Mar 15;9:4619. doi: 10.1038/s41598-019-40549-y

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© The Author(s) 2019

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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Fear conditioning and extinction decrease and increase somatic activities of layer 5 pyramidal neurons in the motor cortex respectively. (A) Representative images of calcium activities of layer 5 pyramidal neurons in GCaMP-expressing transgenic mice. The blue and red triangles indicate somas showing reduced and increased activities to tone respectively as compared to the pre-tone period. (B) The response of the labeled soma in (A, blue) with reduced activity to tone. Changes in the somatic calcium level were measured by ΔF/F₀ (left) and z-score transformation (right). Yellow bars represent tone presentation. (C) The response of the labeled soma in (A, red) with increased activity to tone. Changes in somatic calcium level were measured by ΔF/F₀ (left) and z-score transformation (right). Yellow bars represent tone presentation. (D) Left panel: schematic of experimental design. Somatic calcium activities of layer 5 pyramidal neurons were imaged with or without tone presentation. Right panel: the majority of layer 5 pyramidal neurons exhibited increased somatic calcium activities in response to tone compared to the period of pre-tone (n = 84 somas from 3 mice). (E) Left panel: schematic of experimental design. 2 days after CS-US pairings, somatic calcium activities of layer 5 pyramidal neurons were imaged with or without tone presentation. Right panel: the majority of layer 5 pyramidal neurons exhibited reduced somatic calcium activities in response to CS when compared to the period of pre-CS (n = 85 somas from 4 mice). (F) Left panel: schematic of experimental design. Mice were fear conditioned by CS-US pairings and then subjected to CS extinction. 2 days after CS extinction, somatic calcium activities of layer 5 pyramidal neurons were imaged with or without tone presentation. Right panel: a small percentage of layer 5 pyramidal neurons exhibited reduced somatic calcium activities in response to CS when compared to the period of pre-CS (n = 60 somas from 4 mice).