机构：[1]Department of Biomedicine, University Hospital Basel & University Basel, Hebelstr.20, 4031, Basel, Switzerland[2]Department of Ophthalmology, University Hospital Basel & University Basel, Mittlere Str.91, 4056, Basel, Switzerland[3]Institute of Molecular and Clinical Ophthalmology, Mittlere Str.91, 4056, Basel, Switzerland[4]Department of Ophthalmology, Kantonsspital Aarau, Herzogstrasse 15, 5000, Aarau, Switzerland[5]Beijing Institute of Ophthalmology, Beijing Tongren Hospital, Capital Medical University, Beijing, China首都医科大学附属北京同仁医院研究所眼科研究所临床科室眼科斜视与小儿眼科
The meninges not only surround the brain and the spinal cord but also the optic nerve. Meningeal-derived extracellular matrix (ECM) is a crucial component of the pial basement membrane, glia limitans and important for maintenance of optic nerve axon integrity, homeostasis and retinal ganglion cell health. To get closer insight into optic nerve meningeal-derived ECM composition, we performed proteomic analysis of the sheep optic nerve subarachnoid space (SAS). Candidate components were confirmed in cultures of primary human meningothelial cells (phMECs) and human optic nerve samples. Sheep optic nerve SAS samples were analysed by LC-MS, identified proteins were matched to their human orthologs and filtered using gene lists representing all major ECM components. To validate these findings digital droplet PCR (ddPCR) to evaluate mRNA expression of all candidate components identified was performed on cultures of phMECs. In addition, one protein per major ECM group was stained on human optic nerve sections and on phMEC cultures. Employing LC-MS, 1273 proteins were identified and subjected to bioinformatic analysis. Gene ontology analysis revealed six out of forty-four collagen types (1A1, 1A2, 3A1, 6A2, 6A3 and 14A1), three out of eleven laminin subunits (A4, B2, C1) and six out of twenty-seven hyaluronan binding proteins (CD44, versican (VCAN), Clq binding protein, neurocan (NCAN), brevican (BCAN) and hyalaluronan proteoglycan link protein 2 (HAPLN2)) were present in our cohort. DdPCR in phMEC cell culture confirmed presence of all candidate components except NCAN, BCAN and HAPLN2. Immunohistochemistry (IHC) staining on human optic nerve sections and immunofluorescence (IF) staining on in vitro cultured phMECs showed strong immunopositivity for collagen-typeR alpha 1 (COL1A1), laminin gamma 1 (LAMC1), and VCAN. Fibronectin (FN1) was exclusively present in cultures of phMECs. Using a combined bioinformatics and immunohistological approach, we describe the ECM composition of the optic nerve subarachnoid space. As this space plays an important role in maintaining optic nerve function, a better understanding of ECM composition in this delicate environment might be key to further pathophysiological insight into optic nerve degeneration and associated disorders.
基金：
Messerli-Stiftung (S¨orenberg, Switzerland) and Forschungsfonds Kantonsspital Aarau (Aarau, Switzerland).
第一作者机构：[1]Department of Biomedicine, University Hospital Basel & University Basel, Hebelstr.20, 4031, Basel, Switzerland[5]Beijing Institute of Ophthalmology, Beijing Tongren Hospital, Capital Medical University, Beijing, China
共同第一作者：
通讯作者：
通讯机构：[1]Department of Biomedicine, University Hospital Basel & University Basel, Hebelstr.20, 4031, Basel, Switzerland[2]Department of Ophthalmology, University Hospital Basel & University Basel, Mittlere Str.91, 4056, Basel, Switzerland[*1]University Hospital Basel, Department of Biomedicine, Ocular Pharmacology and Physiology, Laboratory 422, Hebelstr. 20, CH-4031, Basel, Switzerland.
推荐引用方式(GB/T 7714)：
Hao Jie,Kohler Corina,van den Dorpel Hubrecht,et al.The extracellular matrix composition of the optic nerve subarachnoid space[J].EXPERIMENTAL EYE RESEARCH.2020,200:doi:10.1016/j.exer.2020.108250.
APA：
Hao, Jie,Kohler, Corina,van den Dorpel, Hubrecht,Scholl, Hendrik P. N.,Meyer, Peter...&Neutzner, Albert.(2020).The extracellular matrix composition of the optic nerve subarachnoid space.EXPERIMENTAL EYE RESEARCH,200,
MLA：
Hao, Jie,et al."The extracellular matrix composition of the optic nerve subarachnoid space".EXPERIMENTAL EYE RESEARCH 200.(2020)
