Ashworth Laboratories, University of Edinburgh, Institute of Evolutionary Biology, Charlotte Auerbach Road, EH9 3FL, Edinburgh, UK.
British Antarctic Survey, Natural Environment Research Council, High Cross, Madingley Road, CB3 0ET, Cambridge, UK.
BMC Genomics. 2021 Jun 10;22(1):437. doi: 10.1186/s12864-021-07751-7.
Biomineralization by molluscs involves regulated deposition of calcium carbonate crystals within a protein framework to produce complex biocomposite structures. Effective biomineralization is a key trait for aquaculture, and animal resilience under future climate change. While many enzymes and structural proteins have been identified from the shell and in mantle tissue, understanding biomieralization is impeded by a lack of fundamental knowledge of the genes and pathways involved. In adult bivalves, shells are secreted by the mantle tissue during growth, maintenance and repair, with the repair process, in particular, amenable to experimental dissection at the transcriptomic level in individual animals.
Gene expression dynamics were explored in the adult blue mussel, Mytilus edulis, during experimentally induced shell repair, using the two valves of each animal as a matched treatment-control pair. Gene expression was assessed using high-resolution RNA-Seq against a de novo assembled database of functionally annotated transcripts. A large number of differentially expressed transcripts were identified in the repair process. Analysis focused on genes encoding proteins and domains identified in shell biology, using a new database of proteins and domains previously implicated in biomineralization in mussels and other molluscs. The genes implicated in repair included many otherwise novel transcripts that encoded proteins with domains found in other shell matrix proteins, as well as genes previously associated with primary shell formation in larvae. Genes with roles in intracellular signalling and maintenance of membrane resting potential were among the loci implicated in the repair process. While haemocytes have been proposed to be actively involved in repair, no evidence was found for this in the M. edulis data.
The shell repair experimental model and a newly developed shell protein domain database efficiently identified transcripts involved in M. edulis shell production. In particular, the matched pair analysis allowed factoring out of much of the inherent high level of variability between individual mussels. This snapshot of the damage repair process identified a large number of genes putatively involved in biomineralization from initial signalling, through calcium mobilization to shell construction, providing many novel transcripts for future in-depth functional analyses.
软体动物的生物矿化涉及到在蛋白质框架内有规律地沉积碳酸钙晶体,从而产生复杂的生物复合材料结构。有效的生物矿化是水产养殖的关键特征,也是动物在未来气候变化下的适应能力。虽然已经从壳和套膜组织中鉴定出许多酶和结构蛋白,但由于缺乏对涉及的基因和途径的基本知识,生物矿化的理解仍然受到阻碍。在成年双壳类动物中,壳是由套膜组织在生长、维持和修复过程中分泌的,特别是修复过程,在个体动物的转录组水平上可以进行实验性剖析。
在实验诱导的壳修复过程中,利用每个动物的两个瓣作为配对的处理对照,对成年贻贝 Mytilus edulis 中的基因表达动态进行了探索。使用针对功能注释转录本从头组装数据库的高分辨率 RNA-Seq 评估基因表达。在修复过程中鉴定出大量差异表达的转录本。分析集中在壳生物学中鉴定出的编码蛋白和结构域的基因上,使用以前在贻贝和其他软体动物中涉及生物矿化的蛋白和结构域的新数据库。与修复相关的基因包括许多其他新颖的转录本,它们编码的蛋白具有在其他壳基质蛋白中发现的结构域,以及以前与幼虫初级壳形成相关的基因。参与细胞内信号传递和维持膜静息电位的基因也在修复过程中被牵涉到。虽然已经提出血细胞在修复过程中积极参与,但在 M. edulis 数据中没有发现这方面的证据。
壳修复实验模型和新开发的壳蛋白结构域数据库有效地鉴定了参与 M. edulis 壳形成的转录本。特别是,配对分析排除了个体贻贝之间固有的高水平变异性的很大一部分。这个损伤修复过程的快照确定了大量可能涉及生物矿化的基因,从最初的信号传递,通过钙动员到壳结构构建,为未来的深入功能分析提供了许多新的转录本。