Faulhauber Ilse, Mohs Heide, Öchsner Irene
Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Arnimallee 22, D-1000, Berlin 33.
Rouxs Arch Dev Biol. 1986 Sep;195(7):409-416. doi: 10.1007/BF00375745.
The distribution of A(M) subunits of lactate dehydrogenase (mainly LDH) in developing muscle, heart, liver, lung, kidney and cartilage tissue of chicken embryos was examined by the indirect fluorescent antibody technique. Antibodies against porcine LDH, purified by affinity chromatography, were used for this purpose. In special areas of newly formed myofibrils in somitic myoblasts fluorescence was already detected after 4 days of incubation, and located at the same place in muscle tissue of all advanced developmental stages examined. During the myotube stage of muscle development staining was also located in the peripheral thickened cytoplasma of the myotubes. The myocardium did not exhibit any fluorescent staining in the developmental stages examined. Endocardium, epicardium and pericardium, however, were fluorescent in young developmental stages. The liver showed fluorescence in 5- to 8-day embryos mainly in the endothelial cells of the blood sinusoids. In 9- to 12-day embryos the bile ducts became fluorescent. In lungs after 9- to 12-day development the epithelium and the surrounding tissues of bronchi exhibited strong immunofluorescence. The mesonephros exhibited faint granular fluorescence in tubule-forming cells and their membranes after 4-9 days of incubation. Advanced developmental stages only exhibited fluorescent blood cells. This latter staining is at least partly due to non-specific reactions of blood cell membranes with FITC-conjugated anti-rabbit IgG. Cartilage is characterized by non-specific fluorescence, but in embryos older than 8 days strong granular fluorescence of chondrocytes and staining of the perichondrium distinguished sections treated with anti-LDH antibodies from control sections reacted only with FITC-conjugated anti-rabbit IgG. In addition, strong fluorescent staining was detectable in certain areas of the 5-day neural tube and faint staining in the mucosa of the intestine from embryos older than 10 days.
采用间接荧光抗体技术检测了鸡胚发育过程中肌肉、心脏、肝脏、肺、肾脏和软骨组织中乳酸脱氢酶（主要是LDH）的A(M)亚基分布。为此使用了经亲和层析纯化的抗猪LDH抗体。在体节肌母细胞中新形成的肌原纤维的特定区域，孵育4天后就已检测到荧光，且在所检查的所有晚期发育阶段的肌肉组织中荧光都位于同一位置。在肌肉发育的肌管阶段，染色也位于肌管外周增厚的细胞质中。在所检查的发育阶段，心肌未呈现任何荧光染色。然而，心内膜、心外膜和心包在发育早期呈荧光。肝脏在5至8天的胚胎中主要在血窦内皮细胞中呈现荧光。在9至12天的胚胎中，胆管开始呈现荧光。在9至12天发育后的肺中，支气管上皮及其周围组织呈现强烈的免疫荧光。中肾在孵育4至9天后，在肾小管形成细胞及其细胞膜中呈现微弱的颗粒状荧光。晚期发育阶段仅呈现荧光血细胞。后一种染色至少部分是由于血细胞细胞膜与异硫氰酸荧光素（FITC）偶联的抗兔IgG的非特异性反应。软骨的特点是非特异性荧光，但在8天以上的胚胎中，软骨细胞强烈的颗粒状荧光和软骨膜染色使用抗LDH抗体处理的切片与仅用FITC偶联的抗兔IgG反应的对照切片区分开来。此外，在5天神经管的某些区域可检测到强荧光染色，在10天以上胚胎的肠黏膜中可检测到微弱染色。
