Dhawan Sangeeta, Gopinathan K P
Department of Microbiology and Cell Biology, Indian Institute of Science, Sir CV Raman Circle, Bangalore 560012, India.
Gene Expr Patterns. 2004 Jan;4(1):15-23. doi: 10.1016/j.modgep.2003.08.001.
Armadillo/beta-catenin, encoded by the segment polarity gene armadillo functions as a downstream effector of canonical Wnt signals. The expression patterns of Bm wnt-1, -Ci and -engrailed, suggested the presence of Wnt-1 network in the middle silkglands of Bombyx mori. To test this, a homologue of armadillo from B. mori (Bm arm) was cloned by PCR using degenerate primers, designed based on the conserved regions of the protein, and characterized. The cloned region harboured one complete and two incomplete Armadillo (Arm) repeat motifs and was highly conserved across species. Genomic Southern analysis suggested Bm arm to be a multi-copy gene. The expression of Bm arm RNA was first detected at stage 6 of embryogenesis, which reached maximum levels at stage 21C and was maintained until larval hatching. The RNA was expressed uniformly in the embryos, whereas the Arm protein was localized in a segmentally reiterated striped pattern, in conformity with its predicted segment polarity nature. Bm arm was also expressed in the entire silkgland and the transition from third to fourth and fifth larval intermoults was accompanied by an increase in the transcript levels. However, the Arm protein was predominantly localized to the middle silkglands, especially the middle and posterior sub-compartments. The silkglands represented a novel expression domain for arm in Bombyx, and the results were consistent with the existence of a canonical Wnt network in the middle silkglands.
由节段极性基因犰狳编码的犰狳/β-连环蛋白作为经典Wnt信号的下游效应物发挥作用。家蚕中肠丝腺中Bm wnt-1、-Ci和-engrailed的表达模式表明存在Wnt-1网络。为了验证这一点,利用基于该蛋白质保守区域设计的简并引物通过PCR克隆了家蚕犰狳的同源物(Bm arm)并进行了表征。克隆区域含有一个完整和两个不完整的犰狳(Arm)重复基序,并且在物种间高度保守。基因组Southern分析表明Bm arm是一个多拷贝基因。Bm arm RNA的表达在胚胎发育的第6阶段首次检测到,在第21C阶段达到最高水平并一直维持到幼虫孵化。RNA在胚胎中均匀表达,而Arm蛋白则以节段重复的条纹模式定位,与其预测的节段极性性质一致。Bm arm也在整个丝腺中表达,并且从第三龄到第四龄和第五龄幼虫蜕皮的转变伴随着转录水平的增加。然而,Arm蛋白主要定位于中肠丝腺,特别是中后亚区室。丝腺代表了家蚕中arm的一个新的表达结构域,结果与中肠丝腺中存在经典Wnt网络一致。