Maunoury R, Robine S, Pringault E, Huet C, Guénet J L, Gaillard J A, Louvard D
Institut Pasteur, Département de Biologie Moléculaire, Paris, France.
EMBO J. 1988 Nov;7(11):3321-9. doi: 10.1002/j.1460-2075.1988.tb03203.x.
Villin is an evolutionarily well conserved, Ca2+ regulated actin-binding protein, and a major structural component of the brush border of specialized absorptive cells. Using paraffin sections and an affinity purified polyclonal anti-villin antibody, we have investigated the early expression of villin during mouse embryogenesis. Villin is first detectable at the early post-implantation stage in visceral endodermal cells at the periphery of the egg cylinder. In this extra embryonic layer, the expression of villin increases and then persists until full term gestation. In the embryo, villin first appears in gut anlage during the axial rotation. Using the same methodology, villin expression is also demonstrated in differentiating embryoid bodies from a teratocarcinoma. Both in extra embryonic and embryonic extracts, villin expression is confirmed by immunoblot and Northern blot analysis which reveal, respectively, a single polypeptide of 93 kd and an mRNA of 3.4 kb in length, two well defined parameters for adult mouse villin gene expression. The results presented here show that paraffin sections allow very sensitive and highly resolutive detection of antigens in early embryogenesis. They provide a detailed developmental profile of villin expression and demonstrate the usefulness of villin as a marker for epithelial cells involved in absorptive processes.
绒毛蛋白是一种在进化上高度保守、受Ca2+调节的肌动蛋白结合蛋白,是特化吸收细胞刷状缘的主要结构成分。我们使用石蜡切片和亲和纯化的多克隆抗绒毛蛋白抗体,研究了绒毛蛋白在小鼠胚胎发育过程中的早期表达。绒毛蛋白最早可在植入后早期的卵圆柱外周的脏内胚层细胞中检测到。在这个胚外层中,绒毛蛋白的表达增加,然后持续到足月妊娠。在胚胎中,绒毛蛋白在轴向旋转期间首次出现在肠原基中。使用相同的方法,在来自畸胎癌的分化胚状体中也证明了绒毛蛋白的表达。在胚外提取物和胚胎提取物中,通过免疫印迹和Northern印迹分析证实了绒毛蛋白的表达,这两种分析分别揭示了一条93kd的单多肽和一条长度为3.4kb的mRNA,这是成年小鼠绒毛蛋白基因表达的两个明确参数。此处给出的结果表明,石蜡切片能够非常灵敏且高分辨率地检测早期胚胎发育中的抗原。它们提供了绒毛蛋白表达的详细发育概况,并证明了绒毛蛋白作为参与吸收过程的上皮细胞标志物的有用性。