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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. 8 — Related to Fig. 1. a, Schematic diagram showing injection strategy. A given downstream target of MOp-ul was injected with either AAVretro-Cre or RVdGL-Cre and MOp-ul was injected with either AAV1-CAG-FLEX-GFP or AAV1-CAG-FLEX-tdTomato to Cre-dependently label the axonal output for each target-defined population. b, Example images of collateral outputs from different MOp-ul projection neuron types. TEa-projecting neurons (first two columns) were found mostly in L5a and collateralized to all cortical targets and striatum, but not to thalamus or brainstem, characteristic of the IT cell class. Interestingly, the striatal projection was predominately ipsilateral, while output to TEa/ECT was bilateral and projections to PERI exhibited a contralateral bias. In contrast, contralateral CP-projecting neurons (third column) also exhibited an IT projection profile, however they were found primarily in L5b (perhaps a result of AAVretro viral tropism), and displayed strong bilateral projections to striatum, but very little projection to TEa, ECT, or PERI regions. Ipsilateral CP-projecting neurons (fourth column) exhibited a similar profile, but also included L5b ET neurons, which project to ipsilateral striatum, as well as thalamus and brainstem regions. Target-defined ET neurons (columns 5-8) broadly collateralized to all other expected targets of this class, except for thalamic-targeting ET cells (column 5, AAVretro-Cre injection in PO labels L5b, but not L6, thalamic projection due to viral tropism) which displayed little or no projection to lateral medulla (e.g. SPVO, last panel in column), while lateral medulla-targeting ET cells (column 8, SPVO) showed little or no projection to thalamus (e.g. PO, fourth panel in column). In addition, all target-defined ET cell populations collateralized to ipsilateral MOs (second row). Lastly, L6 VAL or VM-projecting neurons (columns 9 and 10) co-targeted all other expected thalamic nuclei (PCN, PO, PF) and the reticular thalamic nucleus (RT). No cortical, striatal, or brainstem collaterals were observed, characteristic of the CT cell class. c, Summary of collateral targeting differences for each major cell class. Scale bars, 500 µm (b).