Aragonite crystallization in primary cell cultures of multicellular isolates from a hard coral, Pocillopora damicornis
Aragonite crystallization in primary cell cultures of multicellular isolates from a hard coral, Pocillopora damicornis
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DOI：
10.1073/pnas.211439698
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发表时间：
2001-10-09
影响因子：
11.1
通讯作者：
Ostrander, GK
中科院分区：
文献类型：
--
作者：
Domart-Coulon, IJ;Elbert, DC;Ostrander, GK
海洋珊瑚礁生态系统的基础是碳酸钙的积累，主要是由硬珊瑚（腔肠动物：珊瑚虫：硬珊瑚）的行动。珊瑚礁的表面覆盖着成群的硬珊瑚虫，碳酸钙以文石形态存款下来，形成连续的钙质骨架。硬珊瑚的骨骼组成和结构是有据可查的，然而，钙化的细胞机制知之甚少。关于所涉珊瑚细胞类型的性质或它们在生物钙化作用中的合作，资料很少。我们报告文石结晶在原代细胞培养的硬珊瑚，Pocillopora damicornis。通过体外自发解离分离顶端珊瑚集落碎片的细胞。在不连续Percoll梯度中通过密度分离单个解离的细胞类型。原代细胞培养显示碱性磷酸酶（ALP）活性的短暂增加，在完整的珊瑚观察到的水平。在粘附的多细胞分离培养物中，酶活化之后是文石沉淀。修改介质的离子配方延长了分离株的维持时间，延迟了ALP活化，延迟了文石沉淀。这些结果表明，在体外结晶的文石在珊瑚细胞培养是可能的，并提供了一个创新的方法来研究造礁珊瑚钙化在细胞水平上。
The foundation of marine coral reef ecosystems is calcium carbonate accumulated primarily by the action of hard corals (Coelenterata: Anthozoa: Scleractinia). Colonial hard coral polyps cover the surface of the reef and deposit calcium carbonate as the aragonite polymorph, stabilized into a continuous calcareous skeleton. Scleractinian coral skeleton composition and architecture are well documented; however, the cellular mechanisms of calcification are poorly understood. There is little information on the nature of the coral cell types involved or their cooperation in biocalcification. We report aragonite crystallization in primary cell cultures of a hard coral, Pocillopora damicornis. Cells of apical coral colony fragments were isolated by spontaneous in vitro dissociation. Single dissociated cell types were separated by density in a discontinuous Percoll gradient. Primary cell cultures displayed a transient increase in alkaline phosphatase (ALP) activity, to the level observed in intact corals. In adherent multicellular isolate cultures, enzyme activation was followed by precipitation of aragonite. Modification of the ionic formulation of the medium prolonged maintenance of isolates, delayed ALP activation, and delayed aragonite precipitation. These results demonstrate that in vitro crystallization of aragonite in coral cell cultures is possible, and provides an innovative approach to investigate reef-building coral calcification at the cellular level.