Falcone M, Miyamoto T, Fierro-Renoy F, Macchia E, DeGroot L J
Department of Medicine, University of Chicago, IL.
Eur J Endocrinol. 1994 Jan;130(1):97-106. doi: 10.1530/eje.0.1300097.
We performed an immunohistochemical study on rat brain and liver during fetal and neonatal life using rabbit antipeptide polyclonal antibodies able to recognize each thyroid hormone receptor (TR) isoform. The expression of TR alpha-1, alpha-2 and beta-1 proteins from 14 days of gestation to 21 days after birth was evaluated. Frozen tissues from 14 (F14), 17 (F17) and 21 (F21)-day-old fetuses and from 5 (N5), 16 (N16) and 21 (N21)-day old newborn rats were stained with anti-TR antibodies using an avidin-biotin-peroxidase system. The antipeptide antibodies utilized in the present study were characterized previously: alpha-144 antibody recognizes both TR alpha-1 and alpha-2; alpha-2-431 antibody is specific for TR variant alpha-2, and beta-62 antibody specifically reacts with the TR beta-1 isoform. The expression of TR alpha-1 was deduced by comparing the staining obtained with alpha-144 and alpha-2-431 antibodies. We demonstrated that each TR isoform is expressed in rat brain from 14 days of gestation and that the alpha isoform was predominant in the early stage. The three TR isoforms were expressed in both neural cell nuclei and in glial cell nuclei. As far as the liver is concerned, at F14 the expression of TR isoforms was weaker in hepatocytes when, on the contrary, TR alpha was clearly detected in hematopoietic cells. The expression of TRs in hepatocytes becomes evident later. The data that we obtained, although not quantitative, emphasize the presence of each TR isoform in brain and liver from 14 days of fetal rat life.
我们使用能够识别每种甲状腺激素受体(TR)亚型的兔抗肽多克隆抗体,对胎鼠和新生鼠的脑和肝进行了免疫组织化学研究。评估了从妊娠第14天到出生后21天TRα-1、α-2和β-1蛋白的表达。使用抗生物素蛋白-生物素-过氧化物酶系统,用抗TR抗体对妊娠第14(F14)、17(F17)和21(F21)天的胎鼠以及出生后5(N5)、16(N16)和21(N21)天的新生大鼠的冷冻组织进行染色。本研究中使用的抗肽抗体特性如下:α-144抗体可识别TRα-1和α-2;α-2-431抗体对TR变体α-2具有特异性,β-62抗体与TRβ-1亚型特异性反应。通过比较用α-144和α-2-431抗体获得的染色结果来推断TRα-1的表达。我们证明,每种TR亚型从妊娠第14天起就在大鼠脑中表达,且α亚型在早期占主导。三种TR亚型在神经细胞核和神经胶质细胞核中均有表达。就肝脏而言,在F14时,TR亚型在肝细胞中的表达较弱,相反,在造血细胞中可清晰检测到TRα。TR在肝细胞中的表达后来变得明显。我们获得的数据虽然不是定量的,但强调了从胎鼠生命的第14天起,脑和肝中每种TR亚型的存在。