Immunofluorescence localization of thyroid hormone receptor protein beta 1 and variant alpha 2 in selected tissues: cerebellar Purkinje cells as a model for beta 1 receptor-mediated developmental effects of thyroid hormone in brain.
Immunofluorescence localization of thyroid hormone receptor protein beta 1 and variant alpha 2 in selected tissues: cerebellar Purkinje cells as a model for beta 1 receptor-mediated developmental effects of thyroid hormone in brain.
复制标题
甲状腺激素受体蛋白β1和变体α2在选定组织中的免疫荧光定位：小脑浦肯野细胞作为大脑中甲状腺激素β1受体介导的发育效应的模型。
DOI：
10.1073/pnas.88.9.3887
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发表时间：
1991
影响因子：
11.1
通讯作者：
Oppenheimer,JH
中科院分区：
文献类型：
--
作者：
Strait,KA;Schwartz,HL;Seybold,VS;Ling,NC;Oppenheimer,JH
大鼠c-erbA β 1 mRNA在出生后的前10天内在大脑中升高，与组织三碘甲状腺原氨酸（T3）水平和T3依赖性大脑发育的增加一致。这些数据表明β 1受体可能介导T3效应。而小脑c-erbA β 1 mRNA水平很低。由于小脑发育，包括浦肯野细胞的树突状分支，是一个T3敏感的过程，我们评估了β 1受体蛋白在小脑发育过程中的水平。产生β 1独特肽区的抗血清。通过体外翻译产物的特异性免疫沉淀、肝核提取物中T3结合活性的85%免疫沉淀和组织提取物的Western印迹分析证明了它们的特异性。使用抗β 1抗血清的免疫组织化学研究染色肝细胞核，但不含T3结合活性或β 1 mRNA的睾丸细胞核。在小脑浦肯野细胞中，免疫荧光信号，定位于细胞核，比在肝脏中看到的更强烈，观察。颗粒细胞内也有较弱的阳性信号。因此，我们可以推断，小脑含有显着浓度的β 1受体蛋白，尽管β 1 mRNA含量低。浦肯野细胞核染色强度和免疫沉淀β 1受体结合能力在新生儿期上升。还制备了非T3结合α 2变体蛋白的抗血清，并观察到独特的荧光图案。颗粒细胞核内可见强荧光，浦肯野细胞未见荧光。睾丸中的α 2荧光很高，与该组织中α 2 mRNA的高水平一致。荧光信号似乎主要来源于分裂的精原细胞。我们的研究结果支持了β 1受体在T3诱导的大脑发育中起核心作用的概念，并强烈表明浦肯野细胞是T3的直接靶点。
Rat c-erbA beta 1 mRNA rises in cerebrum during the first 10 days of life, coincident with an increase in tissue triiodothyronine (T3) levels and T3-dependent brain development. These data suggest that the beta 1 receptor may mediate the T3 effect. However, in cerebellum c-erbA beta 1 mRNA levels were very low. Since cerebellar development, including dendritic arborization of Purkinje cells, is a T3-sensitive process, we assessed the levels of the beta 1 receptor protein in cerebellum during development. Antisera to unique peptide regions of beta 1 were raised. Their specificity was demonstrated by specific immunoprecipitation of the in vitro translated product, 85% immunoprecipitation of the T3 binding activity in hepatic nuclear extracts, and Western blot analysis of tissue extracts. Immunohistochemical studies using anti-beta 1 antiserum stained liver nuclei but not testis nuclei, which contain no T3 binding activity or beta 1 mRNA. In cerebellar Purkinje cells, an immunofluorescent signal, localized to the nucleus and more intense than that seen in the liver, was observed. A positive but weaker signal was also present in the granule cells. Thus, we may infer that the cerebellum contains significant concentrations of beta 1 receptor protein despite the low beta 1 mRNA content. Both the intensity of staining in Purkinje cell nuclei and immunoprecipitable beta 1 receptor binding capacity rose in the neonatal period. Antiserum to the non-T3 binding alpha 2 variant protein was also prepared and a distinctive pattern of fluorescence was observed. Strong fluorescence was seen in the nuclei of granule cells, but none was seen in Purkinje cells. The alpha 2 fluorescence in testis was high, consistent with the high levels of alpha 2 mRNA in this tissue. The fluorescent signal appeared to originate primarily in dividing spermatogonia. Our findings support the concept that the beta 1 receptor plays a central role in T3-induced brain development and strongly suggest that the Purkinje cell is a direct target for T3.