Chronotherapy to reinforce circadian rhythms improves poststroke outcomes and glymphatic function in mice
Emma Waight, Yuxi Zhu, Ashley Caudell, Velia S. Vizcarra, Evan Newbold, Michael J. Giannetto, Evalien Duyvestyn, Estephanie Balbuena, Wei Song, Tanzil M. Arefin, Yuki Mori, Maiken Nedergaard, Lauren M. Hablitz
Emma Waight, Yuxi Zhu, Ashley Caudell, Velia S. Vizcarra, Evan Newbold, Michael J. Giannetto, Evalien Duyvestyn, Estephanie Balbuena, Wei Song, Tanzil M. Arefin, Yuki Mori, Maiken Nedergaard, Lauren M. Hablitz
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Research Article Cell biology Neuroscience

Chronotherapy to reinforce circadian rhythms improves poststroke outcomes and glymphatic function in mice

Abstract

Stroke remains a leading cause of morbidity and mortality worldwide, with few effective interventions to promote recovery. Targeting circadian timing and glymphatic function may represent viable therapeutic strategies. Here, we show that the small-molecule clock modulator, KL001; high-dose melatonin; acute light pulses; and active-phase time-restricted feeding were each sufficient to enhance glymphatic function in mice. Moreover, initiating treatment with either KL001 or active-phase time-restricted feeding 3 days after preclinical models of stroke improved motor outcomes, reduced lesion volume, increased glymphatic flow, and lowered poststroke brain cytokine burden. These findings suggest that reinforcing normal daily rhythmicity after stroke can markedly enhance neurological recovery, even when interventions are initiated several days after stroke onset.

Authors

Emma Waight, Yuxi Zhu, Ashley Caudell, Velia S. Vizcarra, Evan Newbold, Michael J. Giannetto, Evalien Duyvestyn, Estephanie Balbuena, Wei Song, Tanzil M. Arefin, Yuki Mori, Maiken Nedergaard, Lauren M. Hablitz

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Figure 3

Chronotherapy improves glymphatic function and stroke recovery.

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Chronotherapy improves glymphatic function and stroke recovery. (A) Sche...
(A) Schematic of experiment for KL001 treatment after photothrombotic (PT) stroke. (B) Representative IHC (scale bar: 1 mm) for box plot of (C) stroke volume, (D) MPI of glymphatic influx, and (E) wire walk velocity. (F) Schematics of anterior-to-posterior coronal sections (white) with individual mouse photothrombotic stroke sites indicated in transparent red for both vehicle (gray) and KL001 (green). The darker the red, the more stroke sites overlapped between mice. (G) Same as in F, but for transient middle cerebral artery occlusion–induced (tMCAO-induced) lesions. (H) Schematic of experiment for KL001 treatment after tMCAO. (I) Representative IHC (scale bar: 1 mm) for (J) box plot of stroke volume. (K) Representative coronal sections of fluorescent tracer after glymphatic influx assay. Areas within dotted yellow boxes are shown at ×2.5 magnification to the right, with perivascular tracer penetration highlighted with yellow arrows. Images are quantified by the box plot in L. (M) Schematic of experiment for restricted feeding after photothrombotic (PT) stroke. (N) Representative IHC (scale bar: 1 mm) for box plot of (O) stroke volume and (P) wire walk velocity. (Q) Same as in F, but for photothrombotic lesions after restricted feeding. Ad libitum (gray), active fed (blue), and sleep fed (orange) groups are indicated by shading. For all IHC, MAP2 (magenta), GFAP (yellow), and DAPI (cyan) 11 days after stroke. IL, ipsilateral; CL, contralateral. Area with dotted white boxes is shown at higher magnification below. Gray shading indicates sham, and blue shading indicates stroke. For all box plots: vehicle (gray), KL001 (green), active feeding (light blue), sleep feeding (orange) mice. Median and quartiles are shown by box-and-whisker plots, with individual mice shown as colored dots. *P < 0.05, **P < 0.01. C and L: unpaired t tests. D and E: 2-way ANOVA with Tukey’s HSD post hoc analysis. J: Mann-Whitney test. O: 1-way ANOVA test with Tukey’s HSD post hoc analysis. P: Kruskal Wallis test and Dunn’s multiple comparisons test. All statistics are shown in Supplemental Table 1.