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Cited by 64Cerebrospinal fluid and hydrocephalusIon Transport and Channel RegulationBarrier Structure and Function Studies

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A real-time in vivo clearance assay for quantification of glymphatic efflux

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Plá, Virginia, Bork, Peter, Harnpramukkul, Aurakoch, Olveda, Genaro, Ladrón-de-Guevara, Antonio, Giannetto, Michael J., Hussain, Rashad, Wang, Wei, Kelley, Douglas H., Hablitz, Lauren M., Nedergaard, Maiken·2022·Cell Reports
DOI 10.1016/j.celrep.2022.111320
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Abstract

Glymphatic fluid transport eliminates metabolic waste from the brain including amyloid-β, yet the methodology for studying efflux remains rudimentary. Here, we develop a method to evaluate glymphatic real-time clearance. Efflux of Direct Blue 53 (DB53, also T-1824 or Evans Blue) injected into the striatum is quantified by imaging the DB53 signal in the vascular compartment, where it is retained due to its high affinity to albumin. The DB53 signal is detectable as early as 15 min after injection and the efflux kinetics are sharply reduced in mice lacking the water channel aquaporin 4 (AQP4). Pharmacokinetic modeling reveal that DB53 efflux is consistent with the existence of two efflux paths, one with fast kinetics (T 1/2 = 50 min) and another with slow kinetics (T 1/2 = 240 min), in wild-type mice. This in vivo methodology will aid in defining the physiological variables that drive efflux, as well as the impact of brain states or disorders on clearance kinetics.

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  • There was excellent agreement between the photometry data and equation (8), with discrepancies at small times possibly due to dye finding its way across the brain via the ventricles.

    “There was excellent agreement between the photometry data and equation (8), with the discrepancies at small times possibly being due to dye finding its way across the brain via the ventricles16.”

    From Brain clearance is reduced during sleep and anesthesia (2024) · cites this paper as [16]

  • A small dye that moves freely in the parenchyma can be used to accurately quantify brain clearance.

    “However, it has recently been shown16 that a small dye which moves freely in the parenchyma can be used to accurately quantify brain clearance (Fig. 2a).”

    From Brain clearance is reduced during sleep and anesthesia (2024) · cites this paper as [16]

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Authors
Plá, V., Bork, P., Harnpramukkul, A., Olveda, G., Ladrón-de-Guevara, A., Giannetto, M. J., Hussain, R., Wang, W., Kelley, D. H., Hablitz, L. M., & Nedergaard, M.
Affiliations
  • University of Rochester Medicine (US)
  • University of Copenhagen (DK)
  • University of Rochester (US)
Corresponding author
Nedergaard, Maiken
ORCID
Venue
Cell Reports
Publisher
Elsevier BV
Published
2022-09
Volume
40
Issue
11
Pages
111320
ISSN
2211-1247 (ISSN-L)
DOI
10.1016/j.celrep.2022.111320
OpenAlex
W4295897908
Cited by
64
References
13
Citations by year
2026: 12 · 2025: 14 · 2024: 29 · 2023: 8 · 2022: 1
FWCI
7.35
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Reports (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by-nc-nd)
Funders
  • National Institute on Aging (R01NS100366, RF1AG057575)
  • Novo Nordisk
  • National Institute of Neurological Disorders and Stroke
  • Army Research Office (MURI W911NF1910280)
  • Dr. Miriam and Sheldon G. Adelson Medical Research Foundation
  • Government of South Australia
  • National Institutes of Health
  • Lundbeckfonden
Topics
  • Health Sciences › Medicine › Clinical Neurology › Cerebrospinal fluid and hydrocephalus
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
Keywords
glymphatic system, brain waste clearance, Evans blue, albumin binding, aquaporin-4, AQP4 knockout mice, striatum, pharmacokinetic modeling, efflux kinetics, amyloid-beta clearance, real-time in vivo imaging
MeSH terms
Glymphatic System, Animals, Biological Transport, Brain, Kinetics, Mice, Aquaporin 4

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