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. 2020 Sep 2;10:14437. doi: 10.1038/s41598-020-71474-0

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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dGRAPHIC can reconstitute GFP from synaptic contact. (A) In utero electroporation was performed at E13.5 to express NT probes (red nuclear label) in cortical layer IV. (B) After the electroporated mice had grown to adult age (about 2 months old), AAV encoding the CT probe (cytosolic mCherry) was stereotaxically injected into the VB. (C) Lower magnification images of a sample brain several weeks after injection. Reconstituted GFP and nucleic mCherry signals were observed in layer IV. Scale bar, 50 μm. (D) Cytosolic mCherry signal was observed in VB neurons, indicating strong expression of the CT probe. Scale bar, 50 μm. (E) Higher-magnification confocal images of layer IV neurons. The reconstituted GFP signal delineates a cortical spiny stellate cell body and dendrites (white arrowheads), which receive input from VB thalamocortical axons. There are also GFP-positive fibers in layer IV, which are processes of other cortical layer IV neurons that received input from VB axons. Scale bar, 100 μm.