Goyer R A, Haust M D, Cherian M G
Department of Pathology, University of Western Ontario, London, Canada.
Placenta. 1992 Jul-Aug;13(4):349-55. doi: 10.1016/0143-4004(92)90059-3.
Cellular localization of metallothionein (MT) in placenta may provide information on its function as a metal binding protein. Rabbit antibodies to rat liver MT cross-reacted with human MT and were used to localize MT in human term placenta by avidin-biotin peroxidase technique. Serial sections (5 microns) were cut from paraffin-embedded placentae obtained at term from five normal women and incubated with rabbit antibodies to MT. Normal rabbit serum was used as a negative control. The slides were incubated with biotinylated swine anti-rabbit IgG (linking antibody) then with avidin-biotin horseradish peroxidase complex and developed with diaminobenzidine in hydrogen peroxide (0.03 per cent) substrate. The optimum staining of MT was obtained at a 1:800 antibody dilution. MT was identified in fetal amniotic cells, syncytial trophoblasts and villous interstitial cells, and in maternal decidual cells. The presence of MT at specific cellular sites suggests that it may regulate the transplacental transport of metals such as zinc, copper and cadmium. Since the level of cadmium is lower and that of zinc and copper higher in fetal than in maternal blood, this may suggest that placental MT may restrict cadmium while enhancing zinc and copper transport.
金属硫蛋白(MT)在胎盘中的细胞定位可能为其作为金属结合蛋白的功能提供信息。抗大鼠肝脏MT的兔抗体与人类MT发生交叉反应,并通过抗生物素蛋白-生物素过氧化物酶技术用于定位足月人胎盘组织中的MT。从五名正常孕妇足月时获得的石蜡包埋胎盘上切取连续切片(5微米),并与抗MT的兔抗体孵育。正常兔血清用作阴性对照。将玻片与生物素化的猪抗兔IgG(连接抗体)孵育,然后与抗生物素蛋白-生物素辣根过氧化物酶复合物孵育,并用二氨基联苯胺在过氧化氢(0.03%)底物中显色。在抗体稀释度为1:800时获得MT的最佳染色效果。在胎儿羊膜细胞、合体滋养层细胞和绒毛间质细胞以及母体蜕膜细胞中鉴定出MT。MT在特定细胞部位的存在表明它可能调节锌、铜和镉等金属的胎盘转运。由于胎儿血液中镉的水平低于母体血液,而锌和铜的水平高于母体血液,这可能表明胎盘MT可能限制镉的转运,同时增强锌和铜的转运。