Knockdown of MLC1 in primary astrocytes causes cell vacuolation: A MLC disease cell model

NEUROBIOLOGY OF DISEASE

卷:43

Article

Lopez de Heredia, Miguel^3,8  Ridder, Margreet C.⁴  Boor, Ilja⁴  Lien, Chun-Fu⁵  Hagemann, Tracy⁶  Messing, Albee⁶  Gorecki, Dariusz C.⁵  Scheper, Gert C.⁴  Martinez, Albert⁷  Nunes, Virginia^2,3,8  van der Knaap, Marjo S.⁴  Estevez, Raul¹ 

[1] Univ Barcelona, Fac Med, Dept Ciencies Fisiol 2, Secc Fisiol, Lhospitalet De Llobregat 08907, Spain

[2] Univ Barcelona, Secc Genet, Dept Ciencies Fisiol 2, Lhospitalet De Llobregat 08907, Spain

[3] ISCIII, Ctr Invest Red Enfermedades Raras CIBERER, Barcelona, Spain

[4] Vrije Univ Amsterdam, Med Ctr, Dept Pediat Child Neurol, Amsterdam, Netherlands

[5] Univ Portsmouth, Inst Biomed & Biomol Sci, Sch Pharm & Biomed Sci, Portsmouth PO1 2UP, Hants, England

[6] Univ Wisconsin, Madison, WI 53706 USA

[7] Inst Res Biomed, Barcelona, Spain

[8] Lab Genet Mol IDIBELL, Barcelona, Spain

关键词: Astrocyte;    Junctions;    Leukodystrophy;    Volume;   

DOI  :  10.1016/j.nbd.2011.03.015

来源: Elsevier

【 摘 要 】

Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare type of leukodystrophy, in the majority of cases caused by mutations in the MLC1 gene. MRI from MLC patients shows diffuse cerebral white matter signal abnormality and swelling, with evidence of increased water content. Histopathology in a MLC patient shows vacuolation of myelin, which causes the cerebral white matter swelling. MLC1 protein is expressed in astrocytic processes that are part of blood- and cerebrospinal fluid-brain barriers. We aimed to create an astrocyte cell model of MLC disease. The characterization of rat astrocyte cultures revealed MLC1 localization in cell-cell contacts, which contains other proteins described typically in tight and adherent junctions. MLC1 localization in these contacts was demonstrated to depend on the actin cytoskeleton: it was not altered when disrupting the microtubule or the GFAP networks. In human tissues, MLC1 and the protein Zonula Occludens 1 (ZO-1), which is linked to the actin cytoskeleton, co-localized by EM immunostaining and were specifically co-immunoprecipitated. To create an MLC cell model, knockdown of MLC1 in primary astrocytes was performed. Reduction of MLC1 expression resulted in the appearance of intracellular vacuoles. This vacuolation was reversed by the co-expression of human MLC1. Re-examination of a human brain biopsy from an MLC patient revealed that vacuoles were also consistently present in astrocytic processes. Thus, vacuolation of astrocytes is also a hallmark of MLC disease. (C) 2011 Elsevier Inc. All rights reserved.

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