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. 2020 Oct 5;10:16459. doi: 10.1038/s41598-020-73177-y

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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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HA regulates excitatory synapse formation. (A) 10 μm-thick cryosections of 90-day-old cortical spheroids were stained for excitatory synapse markers after 24 h of treatment with purified HA (+ HA) or streptomyces hyaluronidase (− HA). Blue: DAPI, Green: HABP (HA), Red: PSD-95 (post-synaptic marker), Grey: vGlut-1 (pre-synaptic marker). Scale bars for top panels of spheroid images are 100 µm, scale bars for bottom ROI panels for spheroid images are 20 µm. (B) Graphs quantifying the area of individual synapse markers following hyaluronan manipulations. n = 30 spheroid slices per treatment. (C) Quantification of the resulting changes in total excitatory synapse area as determined by co-localization of pre- and post-synaptic markers normalized to DAPI. n = 30 spheroid slices per treatment. Solid black dots represent 95th percentile outliers, one asterisk signifies p value < 0.05, two asterisks signify p value < 0.01.