Paschke R, Elisei R, Vassart G, Ludgate M
I.R.I.B.H.N., Brussels, Belgium.
J Endocrinol Invest. 1993 May;16(5):329-32. doi: 10.1007/BF03348847.
The search for autoantigenic targets in thyroid associated ophthalmopathy (TAO) and a possible link with the thyroid continues. Several authors have presented data suggesting that extrathyroidal tissues, e.g. extraocular muscle (EOM), may contain a functional thyrotropin receptor (TSH-R). We have applied Northern blotting and PCR amplification in an attempt to demonstrate the mRNA for the TSH-R in EOM, which although predominantly composed of muscle, may contain a significant proportion of fibroblasts. In Northern blots of poly(A) mRNA of normal human thyroid, EOM and skeletal muscle (SM) probed with a p32 labeled hTSH-R cDNA, transcripts of 4.6 and 4.4 kb were observed only in the thyroid. PCR amplification was performed on thyroid and EOM cDNAs using pairs of primers designed to amplify exons 1-->9, the extracellular domain (ECD), and exon 10, the intracellular domain (ICD). Both the ECD and ICD were amplified from a thyroidal cDNA library but only the ICD in the EOM cDNA library, this probably being due to the inevitable contamination with genomic DNA. These results do not support the hypothesis that the TSH receptor, as such, would be an autoantigen of EOM involved in TAO.
对甲状腺相关眼病(TAO)自身抗原靶点的寻找以及与甲状腺可能存在的联系仍在继续。几位作者提供的数据表明,甲状腺外组织,如眼外肌(EOM),可能含有功能性促甲状腺激素受体(TSH-R)。我们应用Northern印迹法和PCR扩增技术,试图证明EOM中TSH-R的mRNA,EOM虽然主要由肌肉组成,但可能含有相当比例的成纤维细胞。在用p32标记的hTSH-R cDNA探测正常人甲状腺、EOM和骨骼肌(SM)的多聚腺苷酸(poly(A))mRNA的Northern印迹中,仅在甲状腺中观察到4.6和4.4 kb的转录本。使用设计用于扩增外显子1至9(细胞外结构域,ECD)以及外显子10(细胞内结构域,ICD)的引物对,对甲状腺和EOM的cDNA进行PCR扩增。ECD和ICD均从甲状腺cDNA文库中扩增出来,但仅从EOM cDNA文库中扩增出ICD,这可能是由于不可避免地污染了基因组DNA所致。这些结果不支持TSH受体本身会是参与TAO的EOM自身抗原这一假说。