Impairing Otp homeodomain function in oral ectoderm cells affects skeletogenesis in sea urchin embryos
Impairing Otp homeodomain function in oral ectoderm cells affects skeletogenesis in sea urchin embryos
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DOI：
10.1016/s0012-1606(03)00317-8
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发表时间：
2003-10-01
影响因子：
2.7
通讯作者：
Di Bernardo, M
中科院分区：
文献类型：
--
作者：
Cavalieri, V;Spinelli, G;Di Bernardo, M
在海胆胚胎中，骨骼发生是一系列复杂的分子和细胞事件的结果，这些事件协调了形态发生过程。过去和最近的证据有力地表明，骨骼的启动和生长是严格依赖于从口腔外胚层壁发出的信号。如前所述，Orthopedia（OTP），一个含有同源结构域的转录因子，在口腔外胚层细胞的一个小的子集中特异性表达，可能涉及这个信号通路。在这项研究中，我们利用三种不同的策略来解决的问题，是否OTP是一个上游调控子的sketelogenesis，我们描述了显微注射的影响OTP吗啉代取代的反义寡核苷酸和显性负OTP-enrailed mRNA在Paracentrotus lividus胚胎。我们证明，OTP表达的抑制完全废除骨骼合成。相比之下，共注射Otp mRNA和吗啉代反义寡核苷酸特异性地挽救了骨骼生成程序。此外，由孵化酶启动子驱动的Otp-GFP融合基因构建体的局部外胚层表达诱导异位和异常毛刺发生。我们进一步表明，这种同源异型蛋白的间接目标是骨骼发育特异性基因SM 30，其表达是已知的严格控制下的口腔外胚层领土。基于这些结果，我们得出结论，OTP触发外胚层特异性信号，促进骨骼发育。(C)2003爱思唯尔公司All rights reserved.
In the sea urchin embryo skeletogenesis is the result of a complex series of molecular and cellular events that coordinate the morphogenetic process. Past and recent evidence strongly indicate that skeletal initiation and growth are strictly dependent on signals emanating from the oral ectodermal wall. As previously suggested, Orthopedia (Otp), a homeodomain-containing transcription factor specifically expressed in a small subset of oral ectoderm cells, might be implicated in this signalling pathway. In this study, we utilize three different strategies to address the issue of whether Otp is an upstream regulator of sketelogenesis, We describe the effects of microinjection of Otp morpholino-substituted antisense oligonucleotides and dominant-negative Otp-engrailed mRNA in Paracentrotus lividus embryos. We demonstrate that inhibition of Otp expression completely abolishes skeletal synthesis. By contrast, coinjection of Otp mRNA and the morpholino antisense oligonucleotide specifically rescues the skeletogenic program. In addition, localized ectodermal expression of the Otp-GFP fusion gene construct driven by the hatching enzyme promoter, induces ectopic and abnormal spiculogenesis. We further show that an indirect target of this homeoprotein is the skeletogenic specific gene SM30, whose expression is known to be under the strict control of the oral ectoderm territory. Based on these results, we conclude that Otp triggers the ectoderm-specific signal that promotes skeletogenesis. (C) 2003 Elsevier Inc. All rights reserved.