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. 2024 Jun 13;14:13618. doi: 10.1038/s41598-024-64631-2

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© The Author(s) 2024

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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Spatiotemporal mapping reveals increased microglia and cell size that are attenuated by FTY treatment. 64 regions of interest (ROIs) were selected spanning the entire rostral-caudal axis. Iba-1⁺ area per field within each ROI was then quantified and mapped onto the rodent brain in sham, untreated SAH, and SAH-FTY animals at 2- and 7-days following surgery. (A) There was a clear association between location of bleeding and Iba-1⁺ cell responses, although several distant sites also showed increased signal, typically ipsilateral to arterial perforation. At 2 days, Iba-1⁺ was increased in both untreated SAH and SAH-FTY animals compared to shams. At 7 days, SAH-FTY animals displayed a drastic reduction in Iba-1⁺ compared to untreated SAH animals. (B) Increased Iba-1⁺ was observed in the ipsilateral hemisphere of SAH animals, most pronounced on day 7, where Iba-1⁺ was significantly increased in SAH animals compared to shams and attenuated by FTY. Day 2: Two-way ANOVA with main effects of group (p = 0.032), Šídák’s multiple comparisons test for ipsilateral hemisphere: sham vs. SAH p = 0.072, SAH vs. SAH-FTY p = 0.993, for contralateral hemisphere: sham vs. SAH: p = 0.725, SAH vs. SAH-FTY p = 0.875. Day 7: Two-way ANOVA with main effects of group (p = 0.014), Šídák’s multiple comparisons test for ipsilateral hemisphere: sham vs. SAH p = 0.020, SAH vs. SAH-FTY p = 0.025, for contralateral hemisphere: sham vs. SAH: p = 0.821, SAH vs. SAH-FTY p = 0.645). (C) Similar results were observed with Iba-1⁺ cell size, where increased cell size was observed at 2 days in SAH animals compared to shams, but by day 7 SAH-FTY animals had reduced Iba-1⁺ cell size compared to untreated SAH animals. Day 2: Two-way ANOVA with main effects of group (p = 0.026) and hemisphere (p = 0.024), Šídák’s multiple comparisons test for ipsilateral hemisphere: sham vs. SAH p = 0.037, SAH vs. SAH-FTY p = 0.974, for contralateral hemisphere: sham vs. SAH: p = 0.476, SAH vs. SAH-FTY p = 0.770. Day 7: Two-way ANOVA with main effects of group (p = 0.003), Šídák’s multiple comparisons test for ipsilateral hemisphere: sham vs. SAH p = 0.330, SAH vs. SAH-FTY p = 0.013, for contralateral hemisphere: sham vs. SAH: p = 0.996, SAH vs. SAH-FTY p = 0.081. n = 4–6 animals per group. Data shown as mean ± standard error.