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. 2021 Jul 9;12:4220. doi: 10.1038/s41467-021-24469-y

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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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Fig. 7 — a Schematic outlining the timeline for the neurobehavioral testing. Fer-1 (1 mg/kg) is given i.p. once daily for 7 days before CCI and continued until euthanasia or the MWM test. Motor function is evaluated using Rotarod 2 days after CCI. MWM is utilized to examine spatial memory acquisition. MWM consisted of d–f visible platform testing for 3 days (4 trials per day) to assess motor and visual capabilities, followed by h–j hidden platform testing for 5 days (4 trials per day) to assess spatial learning ability. b A two-tailed unpaired Student’s t test and one-way ANOVA plus Tukey’s test revealed motor activity of inured AAV-Prok2-injected mice is improved versus CCI-alone (P = 0.0232). Addition of AAV-shFbxo10 abolishes this effect (P = 0.0222). Fer-1 attenuated the negative effect of AAV-shFbxo10 and enhanced motor function (P = 0.0263). Data are presented as mean values ± SD (n = 8 mice in sham group and 10 mice per group in other groups). c The rotation speed tolerated is not significantly different between groups except between sham + AAV-NC and CCI + AAV-NC. P = 0.0395 versus CCI + AAV-NC group. Data are presented as mean values ± SD (n = 8 mice in sham + AAV-NC group; n = 10 mice in CCI + AAV-NC group). d–f Latency to platform, distance to platform and swimming speed in the visible platform testing. Data are presented as mean values ± SD (n = 8 mice in sham + AAV-NC group; n = 10 mice per group in other groups). g Representative swimming tracks of the mice in all five groups on the 8th day of the MWM task. h–j During the hidden platform testing, time spent to reach the platform (h), swimming distance (i) and swimming speed (j) are recorded. One-way ANOVA followed by Tukey post hoc test for different groups on the same time point are carried out. Among of them, *(red) means CCI + AAV-NC group versus sham + AAV-NC group; *(blue) means CCI + AAV-Prok2 group versus CCI + AAV-NC group; *(green) means CCI + AAV-Prok2 + shFbxo10 group versus CCI + AAV-Prok2 group; *(orange) means CCI + AAV-Prok2 + shFbxo10+Fer-1 group versus CCI + AAV-Prok2 + shFbxo10 group. Mice in CCI group spend more time (P_(19d) < 0.0001, P_(20d) < 0.0001, and P_(21d) < 0.0001) and travel longer distances (P_(19d) < 0.0001, P_(20d) < 0.0001, and P_(21d) < 0.0001) to reach the platform than sham. AAV-Prok2 mice exhibit a decrease in latency (P_(19d) = 0.0199, P_(20d) < 0.0001, and P_(21d) < 0.0001) and distance (P_(19d) = 0.0126_, P_(20d) < 0.0001, and P_(21d) < 0.0001₎ as the training progressed. Mice injected with AAV_-shFbxo10 group exhibits a significant decline in the ability to learn the spatial location of the submerged platform (P_(19d) = 0.0499, P_(20d) < 0.0001 and P_(21d) < 0.0001 for latency; P_(19d) = 0.0409, P_(20d) < 0.0001, and P_(21d) = 0.0004 for distance)_. However, Fer-1 administration significantly reduces the latency and distance spent on searching for the hidden platform despite AAV-shFbxo10 administration (P_(19d) = 0.0009, P_(20d) < 0.0001 and P_(21d) < 0.0001 for latency; P_(19d) = 0.004, P_(20d) = 0.0002, and P_(21d) < 0.0001 for distance). Swimming speed is not different between the groups. *P < 0.05, **P < 0.01, and ***P < 0.001. A two-way ANOVA with repeated measures followed by Tukey post hoc test is used for the whole groups, which reveals group by day interaction effect in latency to platform (F_16,215 = 3.524, P < 0.0001), swimming distance (F_16,215 = 1.998, P = 0.0145) and swimming speed (F_16,215 = 0.6165, P = 0.8702) during hidden test. For MWM analysis, data are presented as mean values ± SD (n = 8 mice in sham + AAV-NC group; n = 10 mice per group in other groups).