Abstract
Pigment epithelium-derived factor (PEDF) is a neurotrophic factor with neuroprotective, antiangiogenic, and antipermeability effects. In the brain, blood–brain barrier (BBB) function is essential for homeostasis. Its impairment plays a crucial role in the pathophysiology of many neurological diseases, including ischemic stroke. We investigated (a) whether PEDF counteracted vascular endothelial growth factor (VEGF)-induced BBB disruption in the mouse brain, (b) the time course and route of BBB permeability and the dynamics of PEDF expression after cerebral ischemia, and (c) whether intraventricular infusion of PEDF ameliorated brain ischemia by reducing BBB impairment. C57Bl6/N mice received intraparenchymal injections of CSF, VEGF, or a combination of VEGF and PEDF. PEDF increased paracellular but not transcellular BBB integrity as indicated by an increase in the tight junction protein claudin-5. In another group of mice undergoing 60-min middle cerebral artery occlusion (MCAO), transcellular BBB permeability (fibrinogen staining in the absence of a loss of claudin-5) increased as early as 6 h after reperfusion. PEDF immunofluorescence increased at 24 h, which paralleled with a decreased paracellular BBB permeability (claudin-5). PEDF after MCAO originated from the blood stream and endogenous pericytes. In the third experiment, the intraventricular infusion of PEDF decreased edema and cell death after MCAO, potentially mediated by the improvement of the paracellular route of BBB permeability (claudin-5) in the absence of an amelioration of Evans Blue extravasation. Together, our data suggest that PEDF improves BBB function after cerebral ischemia by affecting the paracellular but not the transcellular route. However, further quantitative data of the different routes of BBB permeability will be required to validate our findings.
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18 May 2020
The original version of the article unfortunately contained an error in the unit of the protein concentrations under 'Stereotactic Intraparenchymal Injections' subsection in ‘Methods’ section.
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Acknowledgements
We would like to thank Prof. Ulrich Dirnagl and Prof. Peter Vajkoczy for their support and the helpful discussion of the manuscript.
Funding
This work was possible thanks to the financial support from the Departments of Neurosurgery and Experimental Neurology, by the Charité Rahel Hirsch Habilitation Fellowship and Berlin-Brandenburg Center for Regenerative Therapies Flexible Funds (No. BCRTFF2008-17 and BCRTFF2009-38) all granted to Dr. Ana-Luisa Pina. Marietta Zille was supported by a fellowship granted by the Stiftung der Deutschen Wirtschaft (Foundation of German Business).
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ALP initiated the study and provided the original idea, designed all experiments, and performed in part the intraparenchymal experiments, JK developed further ideas on the study and designed the intraparenchymal experiments. AR designed and performed most experiments. AR, RC, and MNK performed the MCAO surgery. AR and MYT performed the ELISA and staining experiments. AR and MZ analyzed the data. AR and MZ performed the statistical analysis and graphical artwork. AR and MZ wrote and edited the paper. AR and MZ contributed equally to the study. All authors discussed the results and commented on the manuscript.
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All animal experiments were performed in accordance with the national and international guidelines for the care and use of laboratory animals (Tierschutzgesetz der Bundesrepublik Deutschland, European directive, as well as GV-SOLAS and FELASA guidelines and recommendations for laboratory animal welfare). The studies were approved by an ethics committee (Landesamt für Gesundheit und Soziales, Berlin, Germany, permit number G 0270/10).
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Riabinska, A., Zille, M., Terzi, M.Y. et al. Pigment Epithelium-Derived Factor Improves Paracellular Blood–Brain Barrier Integrity in the Normal and Ischemic Mouse Brain. Cell Mol Neurobiol 40, 751–764 (2020). https://doi.org/10.1007/s10571-019-00770-9
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DOI: https://doi.org/10.1007/s10571-019-00770-9