IMMUNOCYTOCHEMICAL ANALYSIS OF INTERMEDIATE FILAMENTS IN EMBRYONIC HEART-CELLS WITH MONOCLONAL-ANTIBODIES TO DESMIN
IMMUNOCYTOCHEMICAL ANALYSIS OF INTERMEDIATE FILAMENTS IN EMBRYONIC HEART-CELLS WITH MONOCLONAL-ANTIBODIES TO DESMIN
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DOI：
10.1083/jcb.98.6.2179
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发表时间：
1984-01-01
影响因子：
7.8
通讯作者：
FISCHMAN, DA
中科院分区：
文献类型：
--
作者：
DANTO, SI;FISCHMAN, DA
用鸡砂囊中间丝蛋白（IFP）制备单克隆抗体（McAb）。对两种抗体D3和D 76进行了详细表征。它们特异性结合结蛋白，但识别不同的表位。在成年鸡中，两种单克隆抗体产生相同的免疫荧光染色模式，在冷冻切片中与所有形式的肌肉组织反应，包括血管平滑肌，但不与其他组织类型反应。在分离的骨骼肌原纤维和在纵向冷冻切片的心脏和骨骼肌，结蛋白检测与单克隆抗体在Z-带和肌原纤维之间的纵向取向的丝束。与成人的这些结果相反，在原代培养的胚胎心肌细胞的中间丝（IF）仅被McAb D3修饰，而McAb D 76与这些细胞完全不反应。鸡胚胎发育早期（Hamburger-Hamilton分期17-18）的心脏冰冻切片显示肌细胞区域，用肌球蛋白特异性McAb进行双重免疫荧光鉴定，这些区域未被McAb D 76染色，尽管类似区域被McAb D3染色。McAb D 76与结蛋白在所有成年心肌细胞中反应，但不与所有胚胎心脏细胞反应，表明胚胎和成年心脏IF在免疫学上是不同的，并暗示在心脏发育过程中IF免疫反应性的转换。
Monoclonal antibodies (McAb) were generated against a preparation of intermediate filament proteins (IFP) from adult chicken gizzard. Two antibodies, D3 and D76, were characterized in detail. They bind specifically to desmin but recognize different epitopes. In the adult chicken, both McAbs produced equivalent immunofluorescent staining patterns, reacting in frozen sections with all forms of muscle tissue, including vascular smooth muscle, but with no other tissue types. In isolated skeletal myofibrils and in longitudinal frozen sections of cardiac and skeletal muscle, desmin was detected with both McAbs at the Z-band and in longitudinally-oriented filament bundles between myofibrils. In contrast to these results in the adult, the intermediate filaments (IF) of embryonic cardiac myocytes in primary cultures were decorated only with McAb D3, whereas McAb D76 was completely unreactive with these cells. Frozen sections through the heart at early stages of embryonic chick development (Hamburger-Hamilton stages 17-18) revealed regions of myocytes, identified by double immunofluorescence with myosin-specific McAb, that were unstained with McAb D76 even though similar regions were stained by McAb D3. That McAb D76 reacted with desmin in all adult cardiac myocytes but not with all embryonic heart cells indicates that embryonic and adult cardiac IF are immunologically distinct and implies a conversion in IF immunoreactivity during cardiac development.