Falchuk Kenneth H, Contin Jennifer M, Dziedzic T Scott, Feng Zhongling, French Thayer C, Heffron Gregory J, Montorzi Marcelo
Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Cambridge, MA 02139, USA.
Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):251-6. doi: 10.1073/pnas.012616099.
The determinants of Xenopus laevis embryos that act before their first cell division are mandatory for the formation of mRNas required to establish the dorsal axis. Although their chemical identities are unknown, a number of their properties have long been recognized. One of the determinants is present in the cytoplasm and is sensitive to UV light. Thus, exposing stage 1 embryos to either standard 254-nm or, as shown here, to 366-nm UV light during the 0.3-0.4 time fraction of their first cycle inactivates the cytoplasmic determinant. As a consequence, both types of irradiated embryos fail to express dorsal markers, e.g., goosecoid and chordin, without affecting formation of ventral markers, e.g., Vent-1. The developmental outcome is dorsal axis-deficient morphology. We report here that biliverdin IXalpha, a normal constituent of cytoplasmic yolk platelets, is photo-transformed by irradiation with either 254- or 366-nm UV light and that the transformation triggers the dorsal axis deficiency. When the 254- or 366-nm UV-irradiated embryos, fated to dorsal axis deficiency, are incubated solely with microM amounts of biliverdin, they recover and form the axis. In contrast, incubation with either in vitro photo-transformed biliverdin or biliverdin IXalpha dimethyl ester does not induce recovery. The results define an approach to produce dorsal axis-deficient embryos by photo-transforming its biliverdin by irradiation with 366-nm UV light and identify an unsuspected role for biliverdin IXalpha in X. laevis embryogenesis.
非洲爪蟾胚胎在第一次细胞分裂之前起作用的决定因素对于建立背轴所需的mRNA的形成是必不可少的。尽管它们的化学特性尚不清楚,但它们的一些特性早已为人所知。其中一个决定因素存在于细胞质中,对紫外线敏感。因此,在第1期胚胎第一个周期的0.3 - 0.4时间段内,将其暴露于标准的254纳米紫外线或此处所示的366纳米紫外线,会使细胞质决定因素失活。结果,两种类型的受辐照胚胎都无法表达背侧标记物,如鹅膏菌素和脊索蛋白,而不影响腹侧标记物,如Vent - 1的形成。发育结果是背轴缺陷形态。我们在此报告,细胞质卵黄小板的正常成分胆红素IXα经254纳米或366纳米紫外线照射后会发生光转化,并且这种转化会引发背轴缺陷。当注定会出现背轴缺陷的254纳米或366纳米紫外线照射的胚胎仅与微摩尔量的胆红素一起孵育时,它们会恢复并形成轴。相比之下,用体外光转化的胆红素或胆红素IXα二甲酯孵育不会诱导恢复。这些结果定义了一种通过用366纳米紫外线照射使胆红素发生光转化来产生背轴缺陷胚胎的方法,并确定了胆红素IXα在非洲爪蟾胚胎发生中一个未被怀疑的作用。