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Flow blockage disrupts cilia-driven fluid transport in the epileptic brain

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Abstract

A carpet of ependymal motile cilia lines the brain ventricular system, forming a network of flow channels and barriers that pattern cerebrospinal fluid (CSF) flow at the surface. This CSF transport system is evolutionary conserved, but its physiological function remains unknown. Here we investigated its potential role in epilepsy with studies focused on CDKL5 deficiency disorder (CDD), a neurodevelopmental disorder with early-onset epilepsy refractory to seizure medications and the most common cause of infant epilepsy. CDKL5 is a highly conserved X-linked gene suggesting its function in regulating cilia length and motion in the green alga Chlamydomonas might have implication in the etiology of CDD. Examination of the structure and function of airway motile cilia revealed both the CDD patients and the Cdkl5 knockout mice exhibit cilia lengthening and abnormal cilia motion. Similar defects were observed for brain ventricular cilia in the Cdkl5 knockout mice. Mapping ependymal cilia generated flow in the ventral third ventricle (v3V), a brain region with important physiological functions showed altered patterning of flow. Tracing of cilia-mediated inflow into v3V with fluorescent dye revealed the appearance of a flow barrier at the inlet of v3V in Cdkl5 knockout mice. Analysis of mice with a mutation in another epilepsy-associated kinase, Yes1, showed the same disturbance of cilia motion and flow patterning. The flow barrier was also observed in the Foxj1± and FOXJ1CreERT:Cdkl5y/fl mice, confirming the contribution of ventricular cilia to the flow disturbances. Importantly, mice exhibiting altered cilia-driven flow also showed increased susceptibility to anesthesia-induced seizure-like activity. The cilia-driven flow disturbance arises from altered cilia beating orientation with the disrupted polarity of the cilia anchoring rootlet meshwork. Together these findings indicate motile cilia disturbances have an essential role in CDD-associated seizures and beyond, suggesting cilia regulating kinases may be a therapeutic target for medication-resistant epilepsy.

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Acknowledgements

We thank Dr Pete Lefebvre for advising about the initial idea that CDD might be a motile ciliopathy. We thank Dr Joe Zhou and Dr Chay Kuo for generously providing Cdkl5fl/fl, Cdkl5y/fl and FOXJ1CreERT mice, and supporting with advice.

Funding

German Research Foundation grant FA 1457/1-1 (RJF); National Institutes of Health grant NIH HL142788 (CWL); National Institutes of Health grant NIH HL132024-01 (CWL); National Institutes of Health grant NIH GM051293 (SMK); MKG’s effort is supported by Department of Biomedical Informatics of University of Pittsburgh School of Medicine.

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Authors and Affiliations

  1. Department of Developmental Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15201, USA

    Regina J. Faubel, Veronica S. Santos Canellas, Tim N. Feinstein & Cecilia W. Lo

  2. Division of Child Neurology, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15201, USA

    Jenna Gaesser

  3. Division of Radiology, University of Pittsburgh, Pittsburgh, PA, 15260, USA

    Nancy H. Beluk

  4. Laboratory for Fluid Physics, Pattern Formation and Biocomplexity, Max Planck Institute for Dynamics and Self-Organization, Am Faßberg 17, 37077, Göttingen, Germany

    Yong Wang

  5. Department of Molecular Biology and Biophysics, And Electron Microscopy Facility, University of Connecticut Health Center, Farmington, CT, 06030-3305, USA

    Maya Yankova & Stephen M. King

  6. Supercomputer Education and Research Centre, Indian Institute of Science, Bangalore, 560012, India

    Kalyani B. Karunakaran

  7. Department of Biomedical Informatics, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15201, USA

    Madhavi K. Ganapathiraju

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  1. Regina J. Faubel
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Contributions

Conceptualization: CWL, RJF; Methodology: CWL, RJF, SMK, YW, MKG; Data collection: RJF, MY; Recruitment of CDD patients: JG, NHB; Data analysis: RJF, VSC, MY; Computational Data analysis: RJF, YW, KBK; Visualization: RJF, TNF, YW, MKG, CWL; Funding acquisition: CWL, SMK, RJF; Project administration: CWL; Supervision: CWL, RJF; Writing—original draft: CWL, RJF; Writing—review and editing: CWL, RJF, SMK, YW, KBK, MKG.

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Correspondence to Cecilia W. Lo.

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Faubel, R.J., Santos Canellas, V.S., Gaesser, J. et al. Flow blockage disrupts cilia-driven fluid transport in the epileptic brain. Acta Neuropathol 144, 691–706 (2022). https://doi.org/10.1007/s00401-022-02463-y

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