Coexpression of two mesenchymal cadherins, cadherin 11 and N-cadherin, on murine fibroblast-like synoviocytes
Coexpression of two mesenchymal cadherins, cadherin 11 and N-cadherin, on murine fibroblast-like synoviocytes
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DOI：
10.1002/art.23369
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发表时间：
2008-04-01
影响因子：
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通讯作者：
Brenner, Michael B.
中科院分区：
文献类型：
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作者：
Agarwal, Sandeep K.;Lee, David M.;Brenner, Michael B.
目标。最近在成纤维细胞样滑膜细胞（FLS）上发现了钙粘蛋白11，并且在小鼠身上的研究已经证明了它在滑膜衬里结构和炎症中的重要性。然而，许多组织表达1个以上的钙粘蛋白。鉴于新近认识到钙粘蛋白在滑膜中的功能意义，本研究旨在确定小鼠FLS是否表达除钙粘蛋白外的其他钙粘蛋白。采用免疫荧光（IF）、生化和免疫组织学技术检测了野生型和cadherin 11缺失小鼠的FLS中cadherin的表达特征。在培养的野生型小鼠FLS细胞接触点观察到Cadherin 11的表达。然而，尽管缺乏细胞表面的cadherin 11，缺乏cadherin 11的小鼠FLS细胞仍然形成了含有β -catenin的亲密细胞-细胞接触。用抗β -连环蛋白抗体对细胞表面生物素化的FLS进行免疫沉淀，结果表明野生型小鼠FLS中存在2个细胞表面连环蛋白相关蛋白，cadherin 11-null小鼠FLS中存在1个细胞表面连环蛋白相关蛋白。通过生化方法和逆转录聚合酶链反应，这些蛋白被确定为N-cadherin和cadherin 11。小鼠滑膜免疫组织化学分析证实了N-cadherin和cadherin 11在滑膜衬里的表达。IF分析显示N-cadherin和cadherin 11在细胞-细胞接触的相同点上共定位。然而，使用抗n -钙粘蛋白或抗钙粘蛋白11抗体均不能共免疫沉淀这两种钙粘蛋白，这表明这两种钙粘蛋白并不包含在相同的分子复合物中。这些发现表明，FLS同时表达N-cadherin和cadherin 11，并表明这些cadherin不包含在同一个分子复合物中。考虑到它们在调节细胞行为中的重要性，了解这些钙粘蛋白如何单独和协同调节FLS行为对于理解滑膜结构和炎症至关重要。
Objective. Cadherin 11 has recently been identified on fibroblast-like synoviocytes (FLS), and studies in mice have demonstrated its importance in synovial lining architecture and inflammation. However, many tissues express more than 1 cadherin. Given the newly appreciated functional significance of cadherins in the synovium, this study was undertaken to determine whether mouse FLS express other cadherins in addition to cadherin 11.Methods. The characterization of cadherin expression was determined in FLS derived from wild-type and cadherin 11-null mice using immunofluorescence (IF), biochemical, and immunohistologic techniques.Results. Cadherin 11 expression was observed at points of cell-cell contact in cultured wild-type mouse FLS. However, despite the lack of cell surface cadherin 11, cadherin 11-null mouse FLS cells still formed intimate cell-cell contacts that contained beta-catenin. Immunoprecipitation of cell surface biotinylated FLS with anti-beta-catenin antibody demonstrated the presence of 2 cell surface catenin-associated proteins in FLS from wild-type mice and 1 in FLS sample from cadherin 11-null mice. Using biochemical approaches and reverse transcriptase-polymerase chain reaction, these proteins were determined to be N-cadherin and cadherin 11. Expression of both N-cadherin and cadherin 11 in the synovial lining was demonstrated by immunohistochemical analysis of mouse synovium. IF analyses demonstrated colocalization of N-cadherin and cadherin 11 to the same points of cell-cell contact. However, the inability to coimmunoprecipitate both cadherins using either anti-N-cadherin or anti-cadherin 11 antibodies suggests that these cadherins are not contained within the same molecular complexes.Conclusion. These findings demonstrate that FLS express both N-cadherin and cadherin 11, and suggest that these cadherins are not contained within the same molecular complex. Given their importance in modulating cellular behavior, understanding how these cadherins regulate FLS behavior individually and in concert will be critical to understanding synovial architecture and inflammation.