Mathisen P M, Miller L
Department of Biological Sciences, University of Illinois, Chicago 60680.
Mol Cell Biol. 1989 May;9(5):1823-31. doi: 10.1128/mcb.9.5.1823-1831.1989.
We have used in vitro explant cultures of Xenopus laevis skin to investigate the role that the thyroid hormone triiodothyronine (T3) plays in activating the 63-kilodalton (kDa) keratin genes. The activation of these genes in vivo requires two distinct steps, one independent of T3 and one dependent on T3. In this report we have shown that the same two steps are required to fully activate the 63-kDa keratin genes in skin explant cultures, and we have characterized the T3-mediated step in greater detail. Unlike the induction of transcription by T3 or steroid hormones in adult tissues, there was a long latent period of approximately 2 days between the addition of T3 to skin cultures and an increase in concentration of keratin mRNA. While the T3 induction of 63-kDa keratin gene transcription cannot occur until age 48, a short transient exposure of stage 40 skin cultures to T3 resulted in high-level expression of these genes 5 days later, when normal siblings had reached stage 48. This result indicates that T3 induces a stable change in epidermal cells which can be expressed much later, after extensive cell proliferation has occurred in the absence of T3. Once the 63-kDa keratin genes were induced, they were stably expressed, and by the end of metamorphosis T3 had no further effect on their expression. The results suggest that T3 induces constitutive expression of the 63-kDa keratin genes during metamorphosis.
我们利用非洲爪蟾皮肤的体外外植体培养来研究甲状腺激素三碘甲状腺原氨酸(T3)在激活63千道尔顿(kDa)角蛋白基因中所起的作用。这些基因在体内的激活需要两个不同的步骤,一个独立于T3,另一个依赖于T3。在本报告中,我们表明在皮肤外植体培养中完全激活63-kDa角蛋白基因也需要相同的两个步骤,并且我们更详细地描述了T3介导的步骤。与成年组织中T3或类固醇激素诱导转录不同,在皮肤培养物中添加T3与角蛋白mRNA浓度增加之间存在大约2天的长潜伏期。虽然63-kDa角蛋白基因转录的T3诱导直到48期才会发生,但将40期皮肤培养物短暂暴露于T3会导致这些基因在5天后高水平表达,而此时正常的同代个体已达到48期。这一结果表明,T3诱导表皮细胞发生稳定变化,这种变化在没有T3的情况下大量细胞增殖后很久才会表达。一旦诱导了63-kDa角蛋白基因,它们就会稳定表达,并且在变态结束时T3对其表达不再有进一步影响。结果表明,T3在变态过程中诱导了63-kDa角蛋白基因的组成型表达。