Function of subendothelial basement membranes in Blood-Brain Barrier

内皮下基底膜在血脑屏障中的功能

• 批准号: 14370434
• 负责人: NINOMIYA Yoshifumi
• 金额: $ 9.02万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370434/
• 关键词: basement membrane; capillary; endothelial cell; type IV collagen; astrocyte

We report the molecular cloning of a new member of the transmembrane-type immunoglobulin superfamily and designate the encoded protein as limitrin since it selectively localized to glia limitans in mouse brain. Limitrin cDNA was obtained using a subtractive hybridization procedure designed to identify molecules responsible for blood-brain barrier function. Western blots using a limitrin-specific antibody demonstrated that the gene product is expressed significantly in mouse brain and primary murine astrocytes, and is distributed in the plasma membrane. Immunohistochemical studies using confocal and electron microscopy clearly demonstrated highly polarized localization in astroglial endfeet in the perivascular region and under the pia mater in vivo. Limitrin is expressed in spinal cord and many areas of the brain but not in the median eminence or subfornical organ (the circumventricular organs) where the blood-brain barrier is lacking. Disruption of the blood-brain barrier by cold injury resulted in a drastic reduction in limitrin expression. Furthermore, during retrieval from cold injury the increased expression of limitrin in perivascular endfeet correlated with the recovery of angiogenesis in capillaries within the lesion margins. Our results suggest that limitrin is physically and functionally associated with the blood-brain barrier, implying that this protein may be useful as a diagnostic determinant of barrier integrity.

我们报道了跨膜型免疫球蛋白超家族的一个新成员的分子克隆，并将其命名为Limitrin，因为它选择性地定位于小鼠脑内的胶质细胞限制蛋白。使用消减杂交法获得Limitrin基因，目的是鉴定与血脑屏障功能有关的分子。免疫印迹结果表明，该基因产物在小鼠脑内和原代培养的小鼠星形胶质细胞中均有显著表达，并分布在细胞膜上。免疫组织化学共聚焦和电子显微镜研究清楚地表明，在活体内血管周围区域和软脑膜下的星形胶质细胞终末足部有高度极化的定位。Limitrin在脊髓和脑的许多区域表达，但在缺乏血脑屏障的正中隆起或穹隆下器官(脑室周围器官)中不表达。冷损伤破坏血脑屏障，导致Limitrin表达急剧减少。此外，在冷损伤修复过程中，血管周围终足中Limitrin表达的增加与病变边缘内毛细血管新生的恢复有关。我们的结果表明，Limitrin在物理上和功能上与血脑屏障有关，这意味着该蛋白可能是一种有用的屏障完整性的诊断决定因素。

项目成果

Bekku Y et al.: "Molecular cloning of Bral2, a novel brain-specific link protein, and immunohistochemical colocalization with brevican in perineuronal nets."Mot Cell Neurosci. 24・1. 148-159 (2003)

Bekku Y 等人：“Bral2（一种新型脑特异性连接蛋白）的分子克隆，以及与 brevican 在神经元网络中的免疫组织化学共定位。”Mot Cell Neurosci 24・1（2003）。

Chen L: "Distribution of the Collagen IV Isoforms in Human Bruch's Membrane."British J Ophtalmol. 87・2. 212-215 (2003)

Chen L：“人类 Bruch 膜中 IV 型胶原蛋白的分布”，英国 J Ophtalmol，87・2 212-215（2003 年）。

Harvey SJ: "Transfer of α5(IV) collagen chain gene to smooth muscle restores in vivo expression of the α6(IV) chain in a canine model of Alport syndrome"Am J Pathol. 162. 873-885 (2003)

Harvey SJ：“将 α5(IV) 胶原链基因转移至平滑肌可恢复阿尔波特综合征犬模型中 α6(IV) 链的体内表达”Am J Pathol。162. 873-885 (2003)

Oohashi T: "Brall, a Brain-Specific Link Protein, Colocalizing with the Versican V2 Isoform at the Nodes of Ranvier in Developing and Adult Mouse Central Nervous Systems."Mol Cell Neurosci. 19・1. 43-57 (2002)

Oohashi T：“Brall，一种大脑特异性连接蛋白，在发育中和成年小鼠中枢神经系统中与 Ranvier 节点共定位。”Mol Cell Neurosci 19・1（2002）。

Tomono Y: "Epitope-defined monoclonal. antibodies against multi plexin collagens demonstrate that type XV and XVIII collagens are expressed in specialized basement membranes"Cell Struct Function. 27・1. 9-20 (2002)

Tomono Y：“针对多聚蛋白胶原蛋白的表位定义的单克隆抗体证明 XV 型和 XVIII 型胶原蛋白在特殊的基底膜中表达”《细胞结构功能》27・1（2002 年）。