In vivo imaging of the barrier properties of the glia limitans during health and neuroinflammation

Pauline Hélie-Legoupil, Florencia Kloster, Javier Pareja, Mykhailo Vladymyrov, Josephine A. Mapunda, Elisa Bouillet, Yannik Oetiker, Irene Spera, Sara Barcos, Amandine Brenna, Adolfo Odriozola, Alyssa Baert, Christoph Fankhauser, Beat Haenni, Steven T. Proulx, Benoît Zuber, Urban Deutsch and Britta Engelhardt ()
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Pauline Hélie-Legoupil: University of Bern
Florencia Kloster: University of Bern
Javier Pareja: University of Bern
Mykhailo Vladymyrov: University of Bern
Josephine A. Mapunda: University of Bern
Elisa Bouillet: University of Bern
Yannik Oetiker: University of Bern
Irene Spera: University of Bern
Sara Barcos: University of Bern
Amandine Brenna: University of Bern
Adolfo Odriozola: University of Bern
Alyssa Baert: University of Bern
Christoph Fankhauser: University of Bern
Beat Haenni: University of Bern
Steven T. Proulx: University of Bern
Benoît Zuber: University of Bern
Urban Deutsch: University of Bern
Britta Engelhardt: University of Bern

Nature Communications, 2025, vol. 16, issue 1, 1-24

Abstract: Abstract The glia limitans ensheathes the entire central nervous system (CNS) parenchyma towards the outer surfaces and the perivascular spaces and is formed by a subset of astrocytes strategically localized at these outer parenchymal borders. Barrier properties of the glia limitans during health and neuroinflammation are incompletely understood. By developing an aquaporin-4 (Aqp4)-mRuby3 knock-in reporter mouse that allows for in vivo imaging of the superficial and perivascular glia limitans, we here show that the glia limitans forms a barrier for soluble mediators, beads and immune cells. Combining the Aqp4-mRuby3 reporter strain with additional reporter alleles for vascular, leptomeningeal or myeloid cells ensures precise localization of immune cells to CNS border zones versus the CNS parenchyma allowing to assign functional roles in CNS immune surveillance versus neuropathology. Availability of the Aqp4-mRuby3 reporter mouse will further advance our understanding of the active role of the glia limitans in CNS immune privilege.

Date: 2025
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DOI: 10.1038/s41467-025-63945-7

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