BINDING PATTERN OF ANTI-RHODOPSIN MONOCLONAL-ANTIBODIES TO PHOTORECEPTOR CELLS - AN IMMUNOCYTOCHEMICAL STUDY
BINDING PATTERN OF ANTI-RHODOPSIN MONOCLONAL-ANTIBODIES TO PHOTORECEPTOR CELLS - AN IMMUNOCYTOCHEMICAL STUDY
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DOI：
10.1016/s0014-4835(89)80022-3
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发表时间：
1989-12-01
影响因子：
3.4
通讯作者：
HARGRAVE, PA
中科院分区：
文献类型：
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作者：
ROHLICH, P;ADAMUS, G;HARGRAVE, PA
一组具有明确表位特异性的抗视紫质单抗(MAb)已经通过免疫细胞化学进行了评估。大部分Ig G类单抗(23/27)呈阳性反应，但仅有少数Ig M类单抗(2/21)具有应用价值。N-末端区域的单抗染色最强，高尔基体、核周膜、内节质膜和突触区染色逐渐减弱。位于色素上皮底部的吞噬小体被染色；锥体细胞始终为阴性。视紫红质C末端的抗体标记相同的细胞结构(吞噬小体除外)，但在结合模式上表现出多样性。其中许多单抗除了与杆状细胞结合外，还与锥体外节结合，并与红/绿和蓝色锥体呈现不同的结合模式。视紫红质序列340-348的特异性抗体标记不同类型的锥体细胞，表明它们的结合部位不同。发现两个单抗标记连接视紫红质‘’跨膜片段的亲水环序列：结合环5-6的单抗K42-41和结合环2-3的单抗A1-55。在固定的视网膜中，除了C末端和N末端外，至少这两个视紫红质序列区域可用于抗体反应。
A panel of anti-rhodopsin monoclonal antibodies (MAbs) of defined epitope specificity has been evaluated by immunocytochemistry. Most of the IgG class MAbs (23/27) gave positive results, but only a few of the IgM class MAbs (2/21) were useful for this application. MAbs specific to the N-terminal region stained rod outer segments most strongly, with progressively less staining in the Golgi, perikarya, plasma membrane of the inner segment, and synaptic region. Phagosomes located basally in the pigment epithelium were stained; cone cells were consistently negative. Antibodies to the C-terminus of rhodopsin labeled the same cell structures (except for phagosomes) but showed diversity in their binding pattern. Many of these MAbs bound to cone outer segments in addition to rods, and showed different patterns of binding to red/green and blue cones. Antibodies specific for rhodopsin sequence 340-348 labeled different types of cone cells, indicating differences in their binding sites. Two MAbs were found to label hydrophilic loop sequences which connect rhodopsin''s transmembrane segment: MAb K42-41 which binds loop 5-6, and MAb A1-55 which binds loop 2-3. At least these two regions of the rhodopsin sequence in addition to the C- and N-termini, are available for antibody reaction in fixed retina.