Characterization of calcium deposition and shell matrix protein secretion in primary mantle tissue culture from the marine pearl oyster Pinctada fucata
Characterization of calcium deposition and shell matrix protein secretion in primary mantle tissue culture from the marine pearl oyster Pinctada fucata
复制标题
海洋珍珠贝珠母贝原代地幔组织培养物中钙沉积和壳基质蛋白分泌的表征
DOI：
10.1007/s10126-008-9081-1
复制
发表时间：
2008-07-01
影响因子：
3
通讯作者：
Zhang, Rongqing
中科院分区：
文献类型：
--
作者：
Gong, Ningping;Ma, Zhuojun;Zhang, Rongqing
在这项研究中，我们建立并表征了一种长期的海洋珍珠牡蛎（Pinctada fucata）的原代胚膜组织培养，用于体外研究珍珠层的生物矿化。在该培养体系中，套组织细胞的生存期可达2个月。RT-PCR证实组织细胞可表达珍珠质基质蛋白，Western blot检测培养基中可溶壳基质蛋白nacrein。另一方面，开始培养15天后，在地幔外植体和细胞外生区产生了大量钙、碳、氧等主要元素的钙沉积。随着栽培时间的延长，钙沉积的数量和大小逐渐增加，其位置和纳米颗粒结构提示其生物成因。这些钙沉积特别出现在地幔组织培养中，而不是在心脏组织培养中。综上所述，这些结果表明，套组织培养物的功能与体内的套细胞相似。本研究为珍珠层生物矿化在细胞水平的进一步研究提供了可靠的途径。
In this study, we established and characterized a long-term primary mantle tissue culture from the marine pearl oyster Pinctada fucata for in vitro investigation of nacre biomineralization. In this culture system, the viability of mantle tissue cells lasted up to 2 months. The tissue cells were demonstrated to express nacre matrix proteins by RT-PCR, and a soluble shell matrix protein, nacrein, was detected in the culture medium by Western blot analysis. On the other hand, 15 days after initiating culture, a large amount of calcium deposits with major elements, including calcium, carbon, and oxygen, were generated in the mantle explants and cell outgrowth area. The quantity and size of calcium deposits increased with the prolonged cultivation, and their location and nanogranular structure suggested their biogenic origin. These calcium deposits specifically appeared in mantle tissue cultures, but not in heart tissue cultures. Taken together, these results demonstrate that the mantle tissue culture functions similarly to mantle cells in vivo. This study provides a reliable approach for the further investigation on nacre biomineralization at the cellular level.