TY - JOUR
T1 - Spreading depolarizations trigger caveolin-1–dependent endothelial transcytosis
AU - Sadeghian, Homa
AU - Lacoste, Baptiste
AU - Qin, Tao
AU - Toussay, Xavier
AU - Rosa, Roberto
AU - Oka, Fumiaki
AU - Chung, David Y.
AU - Takizawa, Tsubasa
AU - Gu, Chenghua
AU - Ayata, Cenk
N1 - Funding Information:
Funded by a Fellowship Award from the International Headache Society, Scholarship Award from the Frontiers in Headache Research, the American Headache Society (to H.S.); The Mahoney postdoctoral fellowship (to B.L.); Japanese Heart Foundation and Bayer Yakuhin Research Grant Abroad (to F.O.); The National Institute of Neurological Disorders and Stroke at the NIH (R25NS065743; to D.Y.C.); Fidelity Biosciences Research Initiative (to C.G.); and NIH (DP1 NS092473 Pioneer Award; to C.G.). The research of C.G. was also supported, in part, by a Faculty Scholar grant from the Howard Hughes Medical Institute; The National Institute of Neurological Disorders and Stroke at NIH (P01NS055104 and R01NS102969; to C.A.); and Fondation Leducq, Heitman Foundation, and Ellison Foundation (to C.A.).
Funding Information:
Funded by a Fellowship Award from the International Headache Society, Scholarship Award from the Frontiers in Headache Research, the American Headache Society (to H.S.); The Mahoney postdoctoral fellowship (to B.L.); Japanese Heart Foundation and Bayer Yakuhin Research Grant Abroad (to F.O.); The National Institute of Neurological Disorders and Stroke at the NIH (R25NS065743; to D.Y.C.); Fidelity Biosciences Research Initiative (to C.-G.); and NIH (DP1 NS092473 Pioneer Award; to C.G.). The research of C.G. was also supported, in part, by a Faculty Scholar grant from the Howard Hughes Medical Institute; The National Institute of Neurological Disorders and Stroke at NIH (P01NS055104 and R01NS102969; to C.A.); and Fondation Leducq, Heit-man Foundation, and Ellison Foundation (to C.A.).
Publisher Copyright:
© 2018 American Neurological Association
PY - 2018/9
Y1 - 2018/9
N2 - Objective: Cortical spreading depolarizations (CSDs) are intense and ubiquitous depolarization waves relevant for the pathophysiology of migraine and brain injury. CSDs disrupt the blood–brain barrier (BBB), but the mechanisms are unknown. Methods: A total of six CSDs were evoked over 1 hour by topical application of 300 mM of KCl or optogenetically with 470 nm (blue) LED over the right hemisphere in anesthetized mice (C57BL/6 J wild type, Thy1-ChR2-YFP line 18, and cav-1–/–). BBB disruption was assessed by Evans blue (2% EB, 3 ml/kg, intra-arterial) or dextran (200 mg/kg, fluorescein, 70,000 MW, intra-arterial) extravasation in parietotemporal cortex at 3 to 24 hours after CSD. Endothelial cell ultrastructure was examined using transmission electron microscopy 0 to 24 hours after the same CSD protocol in order to assess vesicular trafficking, endothelial tight junctions, and pericyte integrity. Mice were treated with vehicle, isoform nonselective rho-associated kinase (ROCK) inhibitor fasudil (10 mg/kg, intraperitoneally 30 minutes before CSD), or ROCK-2 selective inhibitor KD025 (200 mg/kg, per oral twice-daily for 5 doses before CSD). Results: We show that CSD-induced BBB opening to water and large molecules is mediated by increased endothelial transcytosis starting between 3 and 6 hours and lasting approximately 24 hours. Endothelial tight junctions, pericytes, and basement membrane remain preserved after CSDs. Moreover, we show that CSD-induced BBB disruption is exclusively caveolin-1–dependent and requires rho-kinase 2 activity. Importantly, hyperoxia failed to prevent CSD-induced BBB breakdown, suggesting that the latter is independent of tissue hypoxia. Interpretation: Our data elucidate the mechanisms by which CSDs lead to transient BBB disruption, with diagnostic and therapeutic implications for migraine and brain injury.
AB - Objective: Cortical spreading depolarizations (CSDs) are intense and ubiquitous depolarization waves relevant for the pathophysiology of migraine and brain injury. CSDs disrupt the blood–brain barrier (BBB), but the mechanisms are unknown. Methods: A total of six CSDs were evoked over 1 hour by topical application of 300 mM of KCl or optogenetically with 470 nm (blue) LED over the right hemisphere in anesthetized mice (C57BL/6 J wild type, Thy1-ChR2-YFP line 18, and cav-1–/–). BBB disruption was assessed by Evans blue (2% EB, 3 ml/kg, intra-arterial) or dextran (200 mg/kg, fluorescein, 70,000 MW, intra-arterial) extravasation in parietotemporal cortex at 3 to 24 hours after CSD. Endothelial cell ultrastructure was examined using transmission electron microscopy 0 to 24 hours after the same CSD protocol in order to assess vesicular trafficking, endothelial tight junctions, and pericyte integrity. Mice were treated with vehicle, isoform nonselective rho-associated kinase (ROCK) inhibitor fasudil (10 mg/kg, intraperitoneally 30 minutes before CSD), or ROCK-2 selective inhibitor KD025 (200 mg/kg, per oral twice-daily for 5 doses before CSD). Results: We show that CSD-induced BBB opening to water and large molecules is mediated by increased endothelial transcytosis starting between 3 and 6 hours and lasting approximately 24 hours. Endothelial tight junctions, pericytes, and basement membrane remain preserved after CSDs. Moreover, we show that CSD-induced BBB disruption is exclusively caveolin-1–dependent and requires rho-kinase 2 activity. Importantly, hyperoxia failed to prevent CSD-induced BBB breakdown, suggesting that the latter is independent of tissue hypoxia. Interpretation: Our data elucidate the mechanisms by which CSDs lead to transient BBB disruption, with diagnostic and therapeutic implications for migraine and brain injury.
UR - http://www.scopus.com/inward/record.url?scp=85053838456&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85053838456&partnerID=8YFLogxK
U2 - 10.1002/ana.25298
DO - 10.1002/ana.25298
M3 - Article
C2 - 30014540
AN - SCOPUS:85053838456
SN - 0364-5134
VL - 84
SP - 409
EP - 423
JO - Annals of Neurology
JF - Annals of Neurology
IS - 3
ER -