Kawasaki Hideki, Ote Manabu, Okano Kazuhiro, Shimada Toru, Guo-Xing Quan, Mita Kazuei
Faculty of Agriculture, Utsunomiya University, 350 Mine, Utsunomiya, Tochigi 321-8505, Japan.
Gene. 2004 Dec 8;343(1):133-42. doi: 10.1016/j.gene.2004.08.013.
The present study was conducted to clarify what occurs during the metamorphosis of the imaginal disc in insects. To understand the metamorphosis on a molecular level, the changes in expression profiles in the imaginal disc during metamorphosis were investigated. For this purpose, we constructed cDNA libraries from four different stages of wing discs of Bombyx mori, sequenced about 1000 cDNAs randomly collected from each library, and constructed a database of expressed sequence tags (EST). The morphological changes and expression profiles from EST were compared during those four stages. Microarray analysis was applied to quantify the expression of each gene in each stage in order to confirm whether the expression of the genes identified from EST was induced by 20-hydroxyecdysone (20E) in a stage-specific manner. Wing discs showed dynamic morphogenesis in 4-5 days during the preparatory stage of metamorphosis, which was under the control of an ecdysteroid. Different expressed profiles were observed in each of the four different stages by comparison of each EST clone. These profiles reflected the morphological changes of the Bombyx wing disc during metamorphosis. The results of expression profiles from the four stages suggested that the V4 stage was cell proliferating; W0, proliferating and the beginning of differentiation; W2, morphologically changing; W3, cuticle secreting. Microarray analysis showed the effectiveness of its application on 20E induction of genes in wing discs. The wing disc of B. mori is an exceptionally suitable system for understanding the relationship between morphological changes and the distribution of mRNA.
本研究旨在阐明昆虫成虫盘变态过程中发生了什么。为了在分子水平上理解变态过程,我们研究了成虫盘在变态过程中表达谱的变化。为此,我们从家蚕翅盘的四个不同阶段构建了cDNA文库,从每个文库中随机收集约1000个cDNA进行测序,并构建了一个表达序列标签(EST)数据库。在这四个阶段中比较了EST的形态变化和表达谱。应用微阵列分析来量化每个阶段每个基因的表达,以确认从EST中鉴定出的基因的表达是否以阶段特异性方式由20-羟基蜕皮激素(20E)诱导。在变态准备阶段的4-5天内,翅盘显示出动态形态发生,这受蜕皮甾体的控制。通过比较每个EST克隆,在四个不同阶段中的每个阶段都观察到了不同的表达谱。这些谱反映了家蚕翅盘在变态过程中的形态变化。四个阶段的表达谱结果表明,V4阶段是细胞增殖阶段;W0阶段,细胞增殖并开始分化;W2阶段,形态发生变化;W3阶段,角质层分泌。微阵列分析表明其在家蚕翅盘中对20E诱导基因的应用是有效的。家蚕翅盘是理解形态变化与mRNA分布之间关系的一个特别合适的系统。