Jones J C, Vikstrom K L, Goldman R D
Department of Cell Biology and Anatomy, Northwestern University Medical School, Chicago, Illinois 60611.
J Cell Sci. 1987 Nov;88 ( Pt 4):513-20. doi: 10.1242/jcs.88.4.513.
We have prepared both monoclonal and polyclonal antibody preparations directed against the 160/165 x 10(3) Mr glycoproteins (desmogleins) of bovine tongue epithelial desmosomes. The polyclonal antibody preparation recognizes desmosomes in a number of mouse tissues, e.g. mouse skin, heart, bladder and trachea, as determined by immunofluorescence microscopy. Furthermore, the polyclonal antibodies recognize polypeptide(s), present in the high salt, Triton-insoluble residues ('cytoskeleton preparations') of mouse skin, heart, bladder and trachea, which comigrate with the 160/165 x 10(3) Mr glycoproteins of bovine tongue epithelial desmosomes as determined by 'Western' immunoblotting. Conversely, the monoclonal 160/165 x 10(3) Mr antibody preparation recognizes desmosomes of stratified squamous epithelial tissues but not desmosomes in other tissue types. Moreover, whereas the monoclonal antibodies recognize 160/165 x 10(3) Mr polypeptides in mouse skin cell cytoskeletons they show no immunoreactivity with the cytoskeleton preparations of mouse bladder, trachea and heart following immunoblotting. These results suggest therefore that although there are conserved epitopes of the 160/165 x 10(3) Mr glycoproteins there are also epitopes of these molecules which vary from tissue to tissue. Double label immunofluorescence observations of cryostat sections of mouse skin using the monoclonal antibodies and antibodies directed against desmoplakin, a plaque component of desmosomes, reveal that the monoclonal antibodies do not recognize certain desmosomes in basal cells which are recognized by desmoplakin antibodies. Indeed, double label observations of cryostat sections of mouse skin using the monoclonal antibodies and human autoantibodies which react with hemidesmosomal components suggest that the monoclonal antibodies stain desmosomes located along the apical surfaces of basal cells but fail to recognize desmosomes along the lateral surfaces of these same cells.(ABSTRACT TRUNCATED AT 250 WORDS)
我们已经制备了针对牛舌上皮桥粒中160/165×10³Mr糖蛋白（桥粒芯糖蛋白）的单克隆和多克隆抗体制剂。通过免疫荧光显微镜检查确定，多克隆抗体制剂可识别多种小鼠组织中的桥粒，如小鼠皮肤、心脏、膀胱和气管。此外，多克隆抗体可识别存在于小鼠皮肤、心脏、膀胱和气管的高盐、Triton不溶性残渣（“细胞骨架制剂”）中的多肽，通过“Western”免疫印迹法测定，这些多肽与牛舌上皮桥粒的160/165×10³Mr糖蛋白迁移率相同。相反，单克隆160/165×10³Mr抗体制剂可识别复层鳞状上皮组织中的桥粒，但不能识别其他组织类型中的桥粒。此外，虽然单克隆抗体可识别小鼠皮肤细胞骨架中的160/165×10³Mr多肽，但在免疫印迹后，它们与小鼠膀胱、气管和心脏的细胞骨架制剂没有免疫反应性。因此，这些结果表明，虽然160/165×10³Mr糖蛋白存在保守表位，但这些分子的表位也因组织而异。使用单克隆抗体和针对桥粒斑块成分桥粒斑蛋白的抗体对小鼠皮肤冰冻切片进行双标记免疫荧光观察发现，单克隆抗体不能识别基底细胞中某些被桥粒斑蛋白抗体识别的桥粒。实际上，使用单克隆抗体和与半桥粒成分反应的人自身抗体对小鼠皮肤冰冻切片进行双标记观察表明，单克隆抗体可染色位于基底细胞顶端表面的桥粒，但不能识别同一细胞侧面的桥粒。（摘要截短于250字）
