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. 2021 Nov 30;32(2):139–156. doi: 10.1038/s41422-021-00588-5

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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. 5 — a, b Schematic of rabies virus-based monosynaptic tracing (a) and quantification of input connection strength index (b). RV-ENVA-deltaG-dsRed (RVdG) was injected into the VP 2-week after unilateral injection of AAV₉-EF1α-DIO-his-eGFP-2a-TVA, and AAV₉-EF1α-DIO-RVG into the VP of Gad-Cre and vGlut2-Cre mice. Numbers of labeled presynaptic neurons/numbers of starter neurons were plotted [Two-tailed Student’s t-test, Gad n = 4, vGlut2 n = 4, NAc t(6) = 3.318, P = 0.016] *P < 0.05. c–e CTB retrograde labeling of GABA^VP-VM and vGlut2^VP-VM neurons. CTB555 was injected into the VM and AAV₉-EF1α-DIO-H2B-GFP was injected into the VP of Gad-Cre or vGlut2-Cre mice (c). Representative images of retrograde labeling (d). Percentage of GABA^VP-VM and vGlut2^VP-VM neurons (e) [Two-tailed Student’s t-test, Gad n = 5, vGlut2 n = 3, t(6) = −17.902, P = 0.000002.] ***P < 0.001. f–j Optical fibers were implanted in the VM/VP and VM to exert laser stimulation and record CaMP6s signal simultaneously. Confocal images show ChrimsonR⁺ or ChR2⁺ terminals (Red), GCaMP6s⁺ cell bodies (Green), and TH⁺ neurons (Blue) in the VM. Scale bar: 50 μm. Statistical graph of group average GCaMP responses aligned to the onset of optical stimulation and bar graph of peak response to optical stimulation at 20 Hz. AAV₉-hSyn-FLEX-ChrimsonR-tdTomato or AAV₉-EF1α-DIO-hChR2-mCherry was injected into the NAc and AAV₉-EF1α-DIO-GCaMP6s was infected in the VM of Gad-Cre mice. Calcium activity of VM GABAergic neurons in response to optical stimulation of GABA^NAc-VM or GABA^NAc-VP projection was recorded (f, g). [f Control n = 5, ChrimsonR n = 5, Two-tailed Student’s t-test, t(8) = 7.014, P = 0.000111; g Control n = 5, ChR2 n = 5, Mann-Whitney U test, Z = 2.611, P = 0.0079] **P < 0.01 and ***P < 0.001. AAV₉-hSyn-FLEX-ChrimsonR-tdTomato or AAV₉-EF1α-DIO-hChR2-mCherry was infected into the NAc and AAV₉-TH-FlpO and AAV₉-EF1α-fDIO-GCaMP6s were injected into the VM of D1-Cre mice. Activity of VM DAergic neurons in response to optical stimulation of D1^NAc-VM or D1^NAc-VP projection was recorded (h, i). [Mann-Whitney U test: h Control n = 6, ChrimsonR n = 6, Z = 2.882, P = 0.0022; i Control n = 6, ChR2 n = 6, Z = −2.882, P = 0.0022] **P < 0.01. (j) AAV₉-EF1α-DIO-hChR2-mCherry was injected into the NAc and AAV₉-TH-FlpO and AAV₉-EF1α-fDIO-GCaMP6s were injected into the VM of D2-Cre mice. Activity of VM DAergic neurons in response to optical stimulation of D2^NAc-VP projection was recorded. [Mann-Whitney U test, Control n = 6, ChR2 n = 6, Z = −2.882, P = 0.0022] **P < 0.01.