Calcium biominerals occur in all major animal phyla, and through biomolecular control, exhibit such diverse structures as exoskeletons, shells, bones, teeth and earstones (otoliths). Determining the three-dimensional expression of key biomineral proteins, however, has proven challenging as typical protein identification methods either lose spatial resolution during dissolution of the mineral phase or are costly and limited to two-dimensional expression of high abundance proteins. Here we present a modification of the CLARITY and ACT-PRESTO protocols to visualize and confirm, for the first time, the timing of expression and function of two key regulators of biomineralization.

中文翻译：

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生物矿化基质蛋白的原位3D可视化。

钙生物矿物质存在于所有主要动物门中，并且通过生物分子控制，表现出各种结构，例如外骨骼，贝壳，骨骼，牙齿和耳石（耳石）。然而，确定关键生物矿物质蛋白质的三维表达已证明具有挑战性，因为典型的蛋白质鉴定方法要么在矿物质相溶解期间失去空间分辨率，要么成本高昂，并且仅限于高丰度蛋白质的二维表达。在这里，我们提出了对CLARITY和ACT-PRESTO协议的修改，以首次可视化并确认生物矿化的两个关键调节因子的表达时间和功能。