Yajima Mamiko
Tateyama Marine Laboratory, Marine and Coastal Research Center, Ochanomizu University, 11 Koyatsu, Tateyama, Chiba 294-0301, Japan.
Dev Biol. 2007 Jul 15;307(2):272-81. doi: 10.1016/j.ydbio.2007.04.050. Epub 2007 May 6.
Primary mesenchyme cells (PMCs) are solely responsible for the skeletogenesis during early larval development of the sea urchin, but the cells responsible for late larval and adult skeletal formation are not clear. To investigate the origin of larval and adult skeletogenic cells, I first performed transplantation experiments in Pseudocentrotus depressus and Hemicentrotus pulcherrimus, which have different skeletal phenotypes. When P. depressus PMCs were transplanted into H. pulcherrimus embryos, the donor phenotype was observed only in the early larval stage, whereas when secondary mesenchyme cells (SMCs) were transplanted, the donor phenotype was observed in late and metamorphic larvae. Second, a reporter construct driven by the spicule matrix protein 50 (SM50) promoter was introduced into fertilized eggs and their PMCs/SMCs were transplanted. In the resultant 6-armed pluteus, green fluorescent protein (GFP) expression was observed in both PMC and SMC transplantations, suggesting SMC participation in late skeletogenesis. Third, transplanted PMCs or SMCs tagged with GFP were analyzed by PCR in the transgenic chimeras. As a result, SMCs were detected in both larval and adult stages, but GFP from PMCs was undetectable after metamorphosis. Thus, it appears that SMCs participate in skeletogenesis in late development and that PMCs disappear in the adult sea urchin, suggesting that the skeletogenesis may pass from PMCs to SMCs during the late larval stage.
初级间充质细胞(PMC)在海胆幼体早期发育过程中对骨骼形成负全责,但负责幼体后期和成虫骨骼形成的细胞尚不清楚。为了研究幼体和成体骨骼生成细胞的起源,我首先在具有不同骨骼表型的凹顶伪球海胆和光棘球海胆中进行了移植实验。当将凹顶伪球海胆的PMC移植到光棘球海胆胚胎中时,仅在幼体早期观察到供体表型,而当移植次级间充质细胞(SMC)时,在幼体后期和变态期幼虫中观察到供体表型。其次,将由骨针基质蛋白50(SM50)启动子驱动的报告构建体导入受精卵,并对其PMC/SMC进行移植。在产生的六腕长腕幼虫中,在PMC和SMC移植中均观察到绿色荧光蛋白(GFP)表达,表明SMC参与后期骨骼形成。第三,通过PCR在转基因嵌合体中分析用GFP标记的移植的PMC或SMC。结果,在幼虫期和成虫期均检测到SMC,但变态后未检测到来自PMC的GFP。因此,似乎SMC在发育后期参与骨骼形成,并且PMC在成年海胆中消失,这表明在幼体后期骨骼形成可能从PMC转移到SMC。