Purificaiton, distribution and clinical significance of TSH-receptor in human thyroid.

我们试图生产针对人TSH受体的抗体。使用该抗体，检查了TSH受体的分布和分布变化。首先，我们尝试使用具有高滴度TSH受体抗体的患者血清纯化TSH受体抗体。将含有高滴度TSH受体抗体的血清与猪甲状腺膜组分孵育。结合IgG通过酸性缓冲液释放，释放的IgG用IgG亲和层析进一步纯化。使用这种IgG在人甲状腺滤泡细胞中检查TSH受体的定位。在顶端质膜上观察到阳性染色。唯一的一个乐队被发现在摩尔。wt130，000使用免疫印迹。我们还研究了rasp21产物的超微结构定位，该产物参与GTP对腺苷酸环化酶的调节。rasp21产物定位于细胞膜顶端。用免疫组化法检测了人甲状腺中表皮生长因子（EGF）及其受体。这些证据表明，TSH受体仅限于甲状腺滤泡细胞的顶端质膜。目前已研究了TSH受体在各种甲状腺疾病中的分布变化。

We have attempted to produce an antibody for human TSH-receptor. Using this antibody, distribution and changes in distribution of TSH-receptor was examined. First, we attempted to purify the TSH-receptor antibody using the serum of patients with high titer of TSH-receptor antibody. The serum containing high titer of TSH-receptor antibody and porcine thyroid membrane fraction were incubated. The binding IgG was released by acidic buffer and released IgG was further purified with IgG affinity chromatography. Using this IgG the localization of TSH-receptor was examined in human thyroid follicular cells. The positive staining was observed at the apical plasma membrane. The only one band was found at Mol. wt 130,000 using the immunoblotting. We also examined the ultrastructural localization of ras p21 product, which participates on GTP regulation of adenylate cyclase. Similar finding that ras p21 product was localized at apical cell membrane ws obtained. Epidermal growth factor (EGF) and EGF-receptor were also examined immunohistochemically in human thyroid. These combined evidences strongly suggested that TSH-receptor is restricted at apical plasma membrane of thyroid follicular cells. The changes in distribution of TSH-receptro in various thyroid diseases have been studied at present.