Smith L C, Harrington M G, Britten R J, Davidson E H
Division of Biology, California Institute of Technology, Pasadena 91125.
Dev Biol. 1994 Aug;164(2):463-74. doi: 10.1006/dbio.1994.1216.
Eggs and embryos of the purple sea urchin (Strongylocentrotus purpuratus) contain profilin that is partly supplied from maternal sources and partly produced by the gastrula. The maternal profilin protein content is about 13 microM and it persists in the embryo at least through gastrulation. Transcript quantitation from probe excess titrations show that very few profilin gene transcripts are present in the embryo during cleavage, but that they increase at the onset of gastrulation. By in situ hybridization, the newly synthesized profilin transcripts are localized in mesenchyme cells. Profilin gene expression increases when mesenchyme cells initiate migration and filopodial extension and retraction. We show that there are three isoforms of maternal profilin protein produced from the single copy gene during oogenesis. However, the blastula stage embryo only produces the major isoform, whereas the acidic isoform is produced in the early stages of gastrulation and the basic isoform appears by the end of gastrulation. Based on transcript prevalence and protein production rates, our calculations indicate that the amount of new protein produced in the mesenchyme cells in 12 hr is at maximum < 2% of that supplied from maternal sources. Because of the large amount of maternally supplied profilin present in the egg and embryo, we suggest that it may be used in the cytokinetic processes of cleavage. Alternatively, because of the small amount of embryonically produced profilin, we suggest that it may function in the cytoskeletal shape changes required for filopodial extension and motility in the mesenchyme cells during gastrulation.
紫海胆(强壮柱头虫)的卵和胚胎含有肌动蛋白结合蛋白,其部分来自母体供应,部分由原肠胚产生。母体肌动蛋白结合蛋白的含量约为13微摩尔,并且至少在原肠胚形成期持续存在于胚胎中。通过探针过量滴定进行的转录本定量分析表明,在卵裂期胚胎中存在极少的肌动蛋白结合蛋白基因转录本,但在原肠胚形成开始时它们会增加。通过原位杂交,新合成的肌动蛋白结合蛋白转录本定位于间充质细胞中。当间充质细胞开始迁移以及丝状伪足伸展和收缩时,肌动蛋白结合蛋白基因表达增加。我们表明,在卵子发生过程中,单拷贝基因产生了三种母体肌动蛋白结合蛋白异构体。然而,囊胚期胚胎仅产生主要异构体,而酸性异构体在原肠胚形成的早期产生,碱性异构体在原肠胚形成结束时出现。根据转录本丰度和蛋白质产生速率,我们的计算表明,间充质细胞在12小时内产生的新蛋白量最多占母体来源供应蛋白量的2%。由于卵子和胚胎中存在大量母体供应的肌动蛋白结合蛋白,我们认为它可能用于卵裂的细胞分裂过程。或者,由于胚胎产生的肌动蛋白结合蛋白量很少,我们认为它可能在原肠胚形成期间间充质细胞丝状伪足伸展和运动所需的细胞骨架形状变化中发挥作用。