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. 2018 Oct 9;9:4163. doi: 10.1038/s41467-018-06501-w

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© The Author(s) 2018

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. 5 — dCA2 targets a vCA1 site containing PNs that project to the NAc shell. a AAV-DIO-ChR2-eYFP was injected in dCA2 and the retrograde label CTB-647 was injected into the NAc shell of Amigo2-Cre mice. Approximate location of sections in b, c, d, e, f, and g are indicated (position #1, #2, #3, #4, #5, and #6, respectively). b Site of injection of CTB-647 (magenta). c Dorsal HPC section showing ChR2-eYFP (green) expression in dCA2 PN cell bodies (co-expressing RGS14, white) with no CTB-647 staining (lack of magenta). d Transverse section of intermediate HPC showing dCA2 fibers (labeled with ChR2-eYFP, green) distributed throughout CA3 and CA1 and sparse CTB-647 labeling (magenta). e–g Transverse sections progressing through vHPC showing dCA2 fibers (green) in vCA3, distal vCA1 (vCA1a) and adjacent ventral subiculum. Retrograde CTB-647 labeling of projections to NAc shell is seen in deep distal vCA1 (vCA1a) and adjacent ventral subiculum (magenta). h–j High magnification views of dotted area in e. k Quantification of normalized CTB-647 fluorescence in deep versus superficial PNs cell layers in vCA1c, vCA1b, and vCA1a. Ventral CA1 fluorescence was greater in deep versus superficial layers (n = 9 slices, 3 mice, repeated measures two-way ANOVA: deep/superficial F(1,8) = 55.05, P < 0.0001). Fluorescence was greater in distal (vCA1a) versus more proximal (vCA1b and vCA1c) areas (n = 9 slices, 3 mice, repeated measures two-way ANOVA: proximal-distal F(2,16) = 14.96, P = 0.0002, vCA1a vs vCA1b P = 0.0135, vCA1a vs vCA1c P = 0.0002, vCA1b vs vCA1c P = 0.1404, Tukey’s multiple comparisons test). l Quantification of normalized fluorescence intensity of ChR2-eYFP signal in dCA2 projections in vCA1. Signal was more intense in vCA1a compared to vCA1b and vCA1c (n = 9 slices, 3 mice, repeated measures one-way ANOVA: F(2,16) = 7.058, P = 0.0063, vCA1a vs vCA1b P = 0.0231, vCA1a vs vCA1c P = 0.0083, vCA1b vs vCA1c P = 0.8702, Tukey’s multiple comparisons test). Results in k and l show mean ± s.e.m. *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars, 250 µm