Top

Published in:

Open Access 01-10-2019 | Original Article

The Contractile Apparatus Is Essential for the Integrity of the Blood-Brain Barrier After Experimental Subarachnoid Hemorrhage

Authors: Clara Luh, Sergej Feiler, Katrin Frauenknecht, Simon Meyer, Lubomir T. Lubomirov, Axel Neulen, Serge C. Thal

Published in: Translational Stroke Research | Issue 5/2019

Abstract

Development of vasogenic brain edema is a key event contributing to mortality after subarachnoid hemorrhage (SAH). The precise underlying mechanisms at the neurovascular level that lead to disruption of the blood-brain barrier (BBB) are still unknown. Activation of myosin light chain kinases (MLCK) may result in change of endothelial cell shape and opening of the intercellular gap with subsequent vascular leakage. Male C57Bl6 mice were subjected to endovascular perforation. Brain water content was determined by wet-dry ratio and BBB integrity by Evans-Blue extravasation. The specific MLCK inhibitor ML-7 was administered to the mice to determine the role of the contractile apparatus of the neurovascular unit in determining brain water content, BBB integrity, neurofunctional outcome, brain damage, and survival at 7 days after SAH. Inhibition of MLCK significantly reduced BBB permeability (Evans Blue extravasation − 28%) and significantly decreased edema formation in comparison with controls (− 2%). MLCK-treated mice showed reduced intracranial pressure (− 53%), improved neurological outcome at 24 h and 48 h after SAH, and reduced 7-day mortality. Tight junction proteins claudin-5 and zonula occludens-1 levels were not influenced by ML-7 at 24 h after insult. The effect of ML-7 on pMLC was confirmed in brain endothelial cell culture (bEnd.3 cells) subjected to 4-h oxygen-glucose deprivation. The present study indicates that MLCK contributes to blood-brain barrier dysfunction after SAH by a mechanism that does not involve modulation of tight junction protein levels, but via activation of the contractile apparatus of the endothelial cell skeleton. This underlying mechanism may be a promising target for the treatment of SAH.

Available only for authorised users

Claassen J, Carhuapoma JR, Kreiter KT, Du EY, Connolly ES, Mayer SA. Global cerebral edema after subarachnoid hemorrhage: frequency, predictors, and impact on outcome. Stroke. 2002;33(5):1225–32.CrossRefPubMed

Mocco J, Prickett CS, Komotar RJ, Connolly ES, Mayer SA. Potential mechanisms and clinical significance of global cerebral edema following aneurysmal subarachnoid hemorrhage. Neurosurg Focus. 2007;22(5):E7.CrossRefPubMed

Doczi T. The pathogenetic and prognostic significance of blood-brain barrier damage at the acute stage of aneurysmal subarachnoid haemorrhage. Clinical and experimental studies. Acta Neurochir (Wien). 1985;77(3–4):110–32.CrossRef

Kuhlmann CR, Tamaki R, Gamerdinger M, Lessmann V, Behl C, Kempski OS, et al. Inhibition of the myosin light chain kinase prevents hypoxia-induced blood-brain barrier disruption. J Neurochem. 2007;102(2):501–7. https://doi.org/10.1111/j.1471-4159.2007.04506.x.CrossRefPubMed

Afonso PV, Ozden S, Prevost MC, Schmitt C, Seilhean D, Weksler B, et al. Human blood-brain barrier disruption by retroviral-infected lymphocytes: role of myosin light chain kinase in endothelial tight-junction disorganization. J Immunol. 2007;179(4):2576–83.CrossRefPubMed

Haorah J, Heilman D, Knipe B, Chrastil J, Leibhart J, Ghorpade A, et al. Ethanol-induced activation of myosin light chain kinase leads to dysfunction of tight junctions and blood-brain barrier compromise. Alcohol Clin Exp Res. 2005;29(6):999–1009.CrossRefPubMed

Luh C, Kuhlmann CR, Ackermann B, Timaru-Kast R, Luhmann HJ, Behl C, et al. Inhibition of myosin light chain kinase reduces brain edema formation after traumatic brain injury. J Neurochem. 2010;112(4):1015–25. https://doi.org/10.1111/j.1471-4159.2009.06514.x.CrossRefPubMed

Rossi JL, Todd T, Bazan NG, Belayev L. Inhibition of myosin light-chain kinase attenuates cerebral edema after traumatic brain injury in postnatal mice. J Neurotrauma. 2013;30(19):1672–9. https://doi.org/10.1089/neu.2013.2898.CrossRefPubMedPubMedCentral

Feiler S, Friedrich B, Scholler K, Thal SC, Plesnila N. Standardized induction of subarachnoid hemorrhage in mice by intracranial pressure monitoring. J Neurosci Methods. 2010;190(2):164–70. https://doi.org/10.1016/j.jneumeth.2010.05.005.CrossRefPubMed

10.

Thal SC, Plesnila N. Non-invasive intraoperative monitoring of blood pressure and arterial pCO2 during surgical anesthesia in mice. J Neurosci Methods. 2007;159(2):261–7. https://doi.org/10.1016/j.jneumeth.2006.07.016.CrossRefPubMed

11.

Sebastiani A, Hirnet T, Jahn-Eimermacher A, Thal SC. Comparison of speed-vacuum method and heat-drying method to measure brain water content of small brain samples. J Neurosci Methods. 2017;276:73–8. https://doi.org/10.1016/j.jneumeth.2016.11.012.CrossRefPubMed

12.

Thal SC, Mebmer K, Schmid-Elsaesser R, Zausinger S. Neurological impairment in rats after subarachnoid hemorrhage--a comparison of functional tests. J Neurol Sci. 2008;268(1–2):150–9. https://doi.org/10.1016/j.jns.2007.12.002.CrossRefPubMed

13.

Bederson JB, Pitts LH, Tsuji M, Nishimura MC, Davis RL, Bartkowski H. Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke. 1986;17(3):472–6.CrossRefPubMed

14.

Timaru-Kast R, Herbig EL, Luh C, Engelhard K, Thal SC. Influence of age on cerebral housekeeping gene expression for normalization of quantitative polymerase chain reaction after acute brain injury in mice. J Neurotrauma. 2015;32(22):1777–88. https://doi.org/10.1089/neu.2014.3784.CrossRefPubMed

15.

Faul F, Erdfelder E, Buchner A, Lang AG. Statistical power analyses using G*power 3.1: tests for correlation and regression analyses. Behav Res Methods. 2009;41(4):1149–60. https://doi.org/10.3758/BRM.41.4.1149.CrossRefPubMed

16.

Feiler S, Plesnila N, Thal SC, Zausinger S, Scholler K. Contribution of matrix metalloproteinase-9 to cerebral edema and functional outcome following experimental subarachnoid hemorrhage. Cerebrovasc Dis. 2011;32(3):289–95. https://doi.org/10.1159/000328248.CrossRefPubMed

17.

Scholler K, Feiler S, Anetsberger S, Kim SW, Plesnila N. Contribution of bradykinin receptors to the development of secondary brain damage after experimental subarachnoid hemorrhage. Neurosurgery. 2011;68(4):1118–23. https://doi.org/10.1227/NEU.0b013e31820a0024.CrossRefPubMed

18.

Neulen A, Meyer S, Kramer A, Pantel T, Kosterhon M, Kunzelmann S, et al. Large vessel vasospasm is not associated with cerebral cortical Hypoperfusion in a murine model of subarachnoid hemorrhage. Transl Stroke Res. 2018. https://doi.org/10.1007/s12975-018-0647-6.

19.

Suzuki H, Hasegawa Y, Kanamaru K, Zhang JH. Mechanisms of osteopontin-induced stabilization of blood-brain barrier disruption after subarachnoid hemorrhage in rats. Stroke. 2010;41(8):1783–90. https://doi.org/10.1161/STROKEAHA.110.586537.CrossRefPubMedPubMedCentral

20.

Katoh K, Kano Y, Amano M, Kaibuchi K, Fujiwara K. Stress fiber organization regulated by MLCK and rho-kinase in cultured human fibroblasts. Am J Physiol Cell Physiol. 2001;280(6):C1669–79.CrossRefPubMed

21.

Suzuki Y, Shibuya M, Takayasu M, Asano T, Ikegaki I, Satoh S, et al. Protein kinase activity in canine basilar arteries after subarachnoid hemorrhage. Neurosurgery. 1988;22(6 Pt 1):1028–31.CrossRefPubMed

22.

Bulter WE, Peterson JW, Zervas NT, Morgan KG. Intracellular calcium, myosin light chain phosphorylation, and contractile force in experimental cerebral vasospasm. Neurosurgery. 1996;38(4):781–7 discussion 7-8.CrossRefPubMed

23.

Sato M, Tani E, Fujikawa H, Kaibuchi K. Involvement of rho-kinase-mediated phosphorylation of myosin light chain in enhancement of cerebral vasospasm. Circ Res. 2000;87(3):195–200.CrossRefPubMed

24.

Kokubu K, Tani E, Nakano M, Minami N, Shindo H. Effects of ML-9 on experimental delayed cerebral vasospasm. J Neurosurg. 1989;71(6):916–22. https://doi.org/10.3171/jns.1989.71.6.0916.CrossRefPubMed

25.

Prunell GF, Mathiesen T, Diemer NH, Svendgaard NA. Experimental subarachnoid hemorrhage: subarachnoid blood volume, mortality rate, neuronal death, cerebral blood flow, and perfusion pressure in three different rat models. Neurosurgery. 2003;52(1):165–75.PubMed

26.

Scholler K, Trinkl A, Klopotowski M, Thal SC, Plesnila N, Trabold R, et al. Characterization of microvascular basal lamina damage and blood-brain barrier dysfunction following subarachnoid hemorrhage in rats. Brain Res. 2007;1142:237–46. https://doi.org/10.1016/j.brainres.2007.01.034.CrossRefPubMed

27.

Thal SC, Sporer S, Klopotowski M, Thal SE, Woitzik J, Schmid-Elsaesser R, et al. Brain edema formation and neurological impairment after subarachnoid hemorrhage in rats. Laboratory investigation. J Neurosurg. 2009;111(5):988–94. https://doi.org/10.3171/2009.3.JNS08412. CrossRefPubMed

28.

Xiong Y, Wang C, Shi L, Wang L, Zhou Z, Chen D, et al. Myosin light chain kinase: a potential target for treatment of inflammatory diseases. Front Pharmacol. 2017;8:292. https://doi.org/10.3389/fphar.2017.00292.CrossRefPubMedPubMedCentral

29.

Rossi JL, Ralay Ranaivo H, Patel F, Chrzaszcz M, Venkatesan C, Wainwright MS. Albumin causes increased myosin light chain kinase expression in astrocytes via p38 mitogen-activated protein kinase. J Neurosci Res. 2011;89(6):852–61. https://doi.org/10.1002/jnr.22600.CrossRefPubMedPubMedCentral

30.

Jian X, Szaro BG, Schmidt JT. Myosin light chain kinase: expression in neurons and upregulation during axon regeneration. J Neurobiol. 1996;31(3):379–91.CrossRefPubMed

31.

Xu Y, Hu W, Liu Y, Xu P, Li Z, Wu R, et al. P2Y6 receptor-mediated microglial phagocytosis in radiation-induced brain injury. Mol Neurobiol. 2016;53(6):3552–64. https://doi.org/10.1007/s12035-015-9282-3.CrossRefPubMed

32.

Lin H, Pallone TL, Cao C. Murine vasa recta pericyte chloride conductance is controlled by calcium, depolarization, and kinase activity. Am J Phys Regul Integr Comp Phys. 2010;299(5):R1317–25. https://doi.org/10.1152/ajpregu.00129.2010.CrossRef

Title: The Contractile Apparatus Is Essential for the Integrity of the Blood-Brain Barrier After Experimental Subarachnoid Hemorrhage
Authors: Clara Luh
Sergej Feiler
Katrin Frauenknecht
Simon Meyer
Lubomir T. Lubomirov
Axel Neulen
Serge C. Thal
Publication date: 01-10-2019
Publisher: Springer US
Published in: Translational Stroke Research / Issue 5/2019
Print ISSN: 1868-4483
Electronic ISSN: 1868-601X
DOI: https://doi.org/10.1007/s12975-018-0677-0

Keynote webinar | Spotlight on medication adherence

Springer Medicine

The Contractile Apparatus Is Essential for the Integrity of the Blood-Brain Barrier After Experimental Subarachnoid Hemorrhage

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 5/2019

Preclinical Validation of the Therapeutic Potential of Glasgow Oxygen Level Dependent (GOLD) Technology: a Theranostic for Acute Stroke

Management of De Novo Carotid Stenosis and Postintervention Restenosis—Carotid Endarterectomy Versus Carotid Artery Stenting—a Review of Literature

Retinal Vasculature Reactivity During Flicker Light Provocation, Cardiac Stress and Stroke Risk in Africans: The SABPA Study

Induction of Brain Arteriovenous Malformation Through CRISPR/Cas9-Mediated Somatic Alk1 Gene Mutations in Adult Mice

Soluble Epoxide Hydrolase-Derived Linoleic Acid Oxylipins in Serum Are Associated with Periventricular White Matter Hyperintensities and Vascular Cognitive Impairment

Management of Cerebral Microbleeds in Clinical Practice