The microstructural, mineralogical and transcriptional developments of shell biomineralization of Pinctada maxima. Series GSE14305.

Molluscan larval ontogeny is a highly conserved process typical of 3 principal developmental stages. A characteristic unique to each of these stages is shell design, termed prodissoconch I, prodissoconch II and dissoconch. These shells vary in morphology, mineralogy and microstructure. The discrete temporal transitions in shell biomineralization between these larval stages are utilized in this study to investigate transcriptional involvement in several distinct biomineralization events. Scanning electron microscopy and X-ray diffraction analysis of P. maxima larvae and juveniles collected throughout post-embryonic ontogenesis, document the mineralogy and microstructure of each shelled stage as well as establishing a timeline for transitions in biomineralization. P. maxima larval samples most representative of these biomineralization distinctions and transitions were analyzed for differential gene expression on the microarray platform PmaxArray 1.0. A number of transcripts are reported as differentially expressed in correlation to the mineralization events of P. maxima larval ontogeny. Some of those isolated are known shell matrix genes while others are novel, these are discussed in relation to potential shell formation roles. This interdisciplinary investigation has married the shell developments of P. maxima larval ontogeny with corresponding gene expression profiles, furthering the elucidation of shell biomineralization.Microarray is used to examine the temporal differential expression of transcripts from several bivalve larval development stages including 24hrs post fertilization, 3 days, 17 days, 20 days, 23 days, 26 days, 30 days, 35 days, 40 days. Differential expression profiles for transcripts of all the temporal samples was determined based on comparison to a common reference of unfertilized eggs. Each temporal larval sample included in the study has at least 3 replicate hybridizations. Dye flips have been incorporated in the replicates. A total of 46 microarray hybridizations were performed in this investigation for differential expression analysis.

软体动物幼虫的发育过程是一种高度保守的进程，典型地分为三个主要发育阶段。每个阶段独有的特征为贝壳设计，分别称为原分离壳I、原分离壳II和分离壳。这些贝壳在形态、矿物学和微观结构上存在差异。本研究利用这些幼虫阶段间贝壳生物矿化的离散时间过渡，来探究转录因子在多个不同生物矿化事件中的参与。对在整个胚胎后发育过程中收集的P. maxima幼虫和幼体进行扫描电子显微镜和X射线衍射分析，记录了每个贝壳阶段的矿物学和微观结构，并确立了生物矿化过渡的时间表。在PmaxArray 1.0微阵列平台上对代表这些生物矿化差异和过渡的P. maxima幼虫样本进行分析，以研究差异基因表达。其中一些被分离出的转录本已知为贝壳基质基因，而另一些则是新型基因，这些基因与潜在贝壳形成作用的关系被进行讨论。这项跨学科研究将P. maxima幼虫发育过程中的贝壳发展与其相应的基因表达谱相结合，进一步阐明了贝壳生物矿化的机制。微阵列技术被用于检测多个双壳类幼虫发育阶段（包括受精后24小时、3天、17天、20天、23天、26天、30天、35天、40天）转录本的时空差异表达。基于未受精卵的公共参考，确定了所有时间样本转录本的差异表达谱。每个参与研究的幼虫样本都包含至少3次重复杂交。在重复中加入了染色反转。本次研究中共进行了46次微阵列杂交，以进行差异表达分析。