Verhoelst Carla H J, Darras Veerle M, Doulabi Behrouz Zandieh, Reyns Geert, Kühn Eduard R, Van der Geyten Serge
Laboratory of Comparative Endocrinology, Zoological Institute, KU Leuven, Naamsestraat 61, B-3000 Leuven, Belgium.
Mol Cell Endocrinol. 2004 Feb 12;214(1-2):97-105. doi: 10.1016/j.mce.2003.10.074.
Immunocytochemistry using polyclonal anti-type I deiodinase (D1) led to the localization of D1 protein in the internal granule cells of the cerebellum in 1-day-old chicks, which was confirmed by the presence of in vitro D1 activity. Western blot analysis of hepatic and cerebellar extracts revealed a band of 27 kDa. In hypothyroid embryos D1 was expressed in both the internal and external granule cell layer and the signal diminished with more severe hypothyroidism, which is in agreement with the expected downregulation of D1 activity during hypothyroidism. In accordance with the protein data, hypothyroidism resulted in the downregulation of cerebellar D1 mRNA. Finally, histological stainings confirmed that the lack of staining in the external germinal layer of 1-day-old euthyroid chicks was due to the fact that migration of immature granule cells from the external towards the internal layer was completed at this stage while cell migration was retarded in hypothyroid animals.
使用多克隆抗I型脱碘酶(D1)进行免疫细胞化学分析,结果显示在1日龄雏鸡的小脑内颗粒细胞中定位到了D1蛋白,体外D1活性的存在证实了这一点。对肝脏和小脑提取物进行的蛋白质印迹分析显示有一条27 kDa的条带。在甲状腺功能减退的胚胎中,D1在内颗粒层和外颗粒层均有表达,且随着甲状腺功能减退程度加重信号减弱,这与甲状腺功能减退期间D1活性预期的下调一致。与蛋白质数据相符,甲状腺功能减退导致小脑D1 mRNA下调。最后,组织学染色证实,1日龄甲状腺功能正常雏鸡的外生发层缺乏染色是由于在此阶段未成熟颗粒细胞从外层向内层的迁移已完成,而在甲状腺功能减退的动物中细胞迁移受到了阻碍。