Rinkevich Yuval, Douek Jacob, Haber Omer, Rinkevich Baruch, Reshef Ram
Faculty of Biology, Technion-Israel Institute of Technology, Haifa, Israel.
Dev Biol. 2007 Dec 1;312(1):131-46. doi: 10.1016/j.ydbio.2007.09.005. Epub 2007 Sep 17.
The phenomenon of whole body regeneration (WBR) from minute soma fragments is a rare event in chordates, confined to the subfamily of botryllid ascidians and is poorly understood on the cellular and molecular levels. We assembled a list of 1326 ESTs from subtracted mRNA, at early stages of Botrylloides leachi WBR, and classified them into functional categories. Sixty-seven (15%) ESTs with roles in innate immunity signaling were classified into a broad functional group, a result supported by domain search and RT-PCR reactions. Gene ontology analysis for human homologous to the immune gene category, identified 22 significant entries, of which "peptidase activity" and "protease inhibitor activity", stood out as functioning during WBR. Analyzing expressions of serine protease Bl-TrSP, a representative candidate gene from the "peptidase activity" subgroup, revealed low transcript levels in naïve vasculature with upregulated expression during WBR. This was confirmed by in situ hybridization that further elucidated staining restricted to a circulating population of macrophage cells. Furthermore, Bl-TrSP was localized in regeneration niches within vasculature, in regenerating buds, and in buds, during blastogenesis. Functional inhibition of serine protease activity disrupts early remodeling processes of the vasculature microenvironment and hinders WBR. Comparison of genome-wide transcription of WBR with five other developmental processes in ascidians (including metamorphosis, budding and blastogenesis), revealed a broad conservation of immune signaling expressions, suggesting a ubiquitous route of harnessing immune-related genes within a broader range of tunicate developmental context. This, in turn, may have enabled the high diversity of life history traits represented by urochordate ascidians.
在脊索动物中,微小体细胞片段实现全身再生(WBR)的现象十分罕见,仅存在于葡萄状海鞘亚科,在细胞和分子层面上人们对此了解甚少。我们收集了柄海鞘WBR早期阶段经消减mRNA得到的1326条EST序列,并将它们分类到不同的功能类别中。67条(15%)在先天免疫信号传导中起作用的EST序列被归为一个宽泛的功能组,这一结果得到了结构域搜索和RT-PCR反应的支持。对与免疫基因类别同源的人类基因进行基因本体分析,确定了22个显著条目,其中“肽酶活性”和“蛋白酶抑制剂活性”在WBR过程中发挥作用尤为突出。分析来自“肽酶活性”亚组的代表性候选基因丝氨酸蛋白酶Bl-TrSP的表达情况,发现在未成熟脉管系统中其转录水平较低,而在WBR过程中表达上调。原位杂交证实了这一点,进一步阐明染色仅限于循环的巨噬细胞群体。此外,在芽殖发生过程中,Bl-TrSP定位于脉管系统内的再生龛、再生芽和芽中。丝氨酸蛋白酶活性的功能抑制会破坏脉管系统微环境的早期重塑过程并阻碍WBR。将WBR的全基因组转录与海鞘的其他五个发育过程(包括变态、出芽和芽殖发生)进行比较,发现免疫信号表达具有广泛的保守性,这表明在更广泛的被囊动物发育背景中存在利用免疫相关基因的普遍途径。反过来,这可能促成了尾索动物海鞘所代表的生活史特征的高度多样性。