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.

我们报告了跨膜型免疫球蛋白超家族的新成员的分子克隆，并将编码的蛋白指定为极限蛋白，因为它有选择地将其定位于小鼠大脑中的胶质极限。使用旨在识别负责血脑屏障功能的分子的减法杂交程序获得了极限蛋白cDNA。使用极限特异性抗体的蛋白质印迹表明，基因产物在小鼠脑和原代鼠星形胶质细胞中显着表达，并且分布在质膜中。使用共聚焦和电子显微镜的免疫组织化学研究清楚地表明，在血管周围区域和体内PIA Mater下，星形胶质细胞终极的定位高度极化。极限蛋白在脊髓和大脑的许多区域中表达，但在缺乏血脑屏障的中位数或下型器官（弯曲的器官）中表达。通过冷损伤对血脑屏障的破坏导致极限蛋白表达的急剧降低。此外，在冷损伤中检索期间，血管周终端中极限蛋白的表达增加与病变边缘内毛细血管中血管生成的恢复相关。我们的结果表明，极限蛋白在物理和功能上与血脑屏障相关，这意味着该蛋白可能是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）。

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）。

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)

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 et al.: "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 等人：“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体内 α6(IV) 链的表达。”Am J Pathol. 162. 873-885 (2003) ）