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. 2021 Oct 6;598(7879):159–166. doi: 10.1038/s41586-021-03970-w

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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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Extended Data Fig. 11 — Related to Fig. 2. a, To distinguish fluorescent labeling of synaptic boutons versus axons of passage in a given MOp target, AAV-hSyn-mRuby2-sypEGFP was injected into MOp-ul and the anterior pretectal nucleus (APN) was examined for the presence of synaptophysin-tagged EGFP+ boutons (green) and mRuby2+ axons (red). b, Injection site in MOp-ul. c, Labeling of boutons and axons in APN at 4X, 10X (d), and 40X magnification (e), confirming synaptic innervation of the downstream structure. f, Similarly, AAV1-hSyn-Cre may be used to confirm and quantify synaptic connectivity in a given target region following anterograde transsynaptic spread of the virus to downstream neurons and subsequent expression of tdTomato in Ai14 Cre-reporter mice. g, Injection site in MOp-ul. h, Post-synaptically labeled tdTomato+ cells (red) in APN at 4X, 10X (i), and 40X magnification (j) confirming a similar pattern of innervation as shown in (d). Blue, fluorescent Nissl stain. Values in mm relative to bregma. Scale bars, 1 mm (b, c, g, h), 200 µm (d, i), 50 µm (e, j).