Tong Xiaoling, He Songzhen, Chen Jun, Hu Hai, Xiang Zhonghuai, Lu Cheng, Dai Fangyin
State Key Laboratory of Silkworm Genome Biology, Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing, 400700, China.
Sci Rep. 2015 Jul 27;5:12562. doi: 10.1038/srep12562.
Laminins are important basement membrane (BM) components with crucial roles in development. The numbers of laminin isoforms in various organisms are determined by the composition of the different α, β, and γ chains, and their coding genes, which are variable across spieces. In insects, only two α, one β, and one γ chains have been identified thus far. Here, we isolated a novel laminin β gene, BmLanB1-w, by positional cloning of the mutant (crayfish, cf) with blistered wings in silkworm. Gene structure analysis showed that a 2 bp deletion of the BmLanB1-w gene in the cf mutant caused a frame-shift in the open reading frame (ORF) and generated a premature stop codon. Knockdown of the BmLanB1-w gene produced individuals exhibiting blistered wings, indicating that this laminin gene was required for cell adhesion during wing development. We also identified laminin homologs in different species and showed that two copies of β laminin likely originated in Lepidoptera during evolution. Furthermore, phylogenetic and gene expression analyses of silkworm laminin genes revealed that the BmLanB1-w gene is newly evolved, and is required for wing-specific cell adhesion. This is the first report showing the tissue specific distribution and functional differentiation of β laminin in insects.
层粘连蛋白是重要的基底膜(BM)成分,在发育过程中起关键作用。不同生物体中层粘连蛋白异构体的数量由不同的α、β和γ链及其编码基因的组成决定,这些基因在不同物种间存在差异。在昆虫中,迄今为止仅鉴定出两条α链、一条β链和一条γ链。在此,我们通过对家蚕中翅翼起泡突变体(小龙虾,cf)进行定位克隆,分离出一个新的层粘连蛋白β基因BmLanB1-w。基因结构分析表明,cf突变体中BmLanB1-w基因缺失2个碱基对导致开放阅读框(ORF)发生移码,并产生一个提前终止密码子。敲低BmLanB1-w基因会产生翅翼起泡的个体,这表明该层粘连蛋白基因在翅翼发育过程中对细胞黏附是必需的。我们还鉴定了不同物种中的层粘连蛋白同源物,并表明β层粘连蛋白的两个拷贝可能在进化过程中起源于鳞翅目。此外,对家蚕层粘连蛋白基因的系统发育和基因表达分析表明,BmLanB1-w基因是新进化的,并且是翅翼特异性细胞黏附所必需的。这是首次报道昆虫中β层粘连蛋白的组织特异性分布和功能分化。