Alfei L, Onali A, Spanò L, Colombari P T, Altavista P L, De Vita R
Animal and Human Biology Department, University of Rome La Sapienza, Italy.
Eur J Histochem. 1994;38(2):151-62.
The possibility of detecting in situ proliferating myosatellite cells during postlarval muscle growth in the carp (Cyprinus carpio, L.) by means of BrdU and PCNA (Cyclin) immunohistochemistry has been evaluated on paraffin embedded sections. Nine subadult stages were defined according to the body length and weight. The fish were injected intraperitoneally with BrdU and fixed one hour later. Adjacent cross sections mounted on glass slides were incubated with monoclonal antiBrdU (1:100) and antiPCNA (1:200) antibody. The proliferative rate, defined as the percentage of labelled cells for each stage, was correlated to the corresponding percentage of small fibers (area less than 200 microns 2) determined by morphometric analysis. Desmin expression, immunocytochemically detected, was aimed at discriminating between the postmitotic and stem myosatellite cells. A low myosatellite cell proliferative activity (2-7%) throughout the carp growth period considered was demonstrated. Quantitative analysis provided evidence that during growth stages in which small fibers are numerous, the myosatellite cell proliferative activity is low and it increases when the small fibers number decreases. It is suggested that an age dependence of myosatellite cell proliferative rate controls the recruitment of new fibers in the carp.
通过BrdU和PCNA（细胞周期蛋白）免疫组织化学方法，在石蜡包埋切片上评估了鲤鱼（Cyprinus carpio, L.）幼体后肌肉生长过程中原位增殖的肌卫星细胞的可能性。根据体长和体重定义了九个亚成体阶段。给鱼腹腔注射BrdU，一小时后固定。将相邻的横切片安装在载玻片上，用单克隆抗BrdU（1:100）和抗PCNA（1:200）抗体孵育。增殖率定义为每个阶段标记细胞的百分比，与通过形态计量分析确定的小纤维（面积小于200平方微米）的相应百分比相关。通过免疫细胞化学检测结蛋白表达，旨在区分有丝分裂后的和干细胞性肌卫星细胞。结果表明，在整个所考虑的鲤鱼生长期间，肌卫星细胞增殖活性较低（2%-7%）。定量分析提供的证据表明，在小纤维数量众多的生长阶段，肌卫星细胞增殖活性较低，而当小纤维数量减少时，其增殖活性增加。研究表明，肌卫星细胞增殖率的年龄依赖性控制着鲤鱼新纤维的募集。
