Demineralization and sectioning of human kidney stones: A molecular investigation revealing the spatial heterogeneity of the stone matrix.
Demineralization and sectioning of human kidney stones: A molecular investigation revealing the spatial heterogeneity of the stone matrix.
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DOI：
10.14814/phy2.14658
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发表时间：
2021-01
影响因子：
2.5
通讯作者：
Williams JC Jr
中科院分区：
文献类型：
--
作者：
Canela VH;Bledsoe SB;Lingeman JE;Gerber G;Worcester EM;El-Achkar TM;Williams JC Jr
肾结石形成的分子机制在很大程度上是未知的。尿液中的有机分子联合收割机与矿物晶体结合形成结石，但对结石基质的分析显示，有一千多种不同的蛋白质，没有线索表明哪些对结石生长很重要。存在于石头的每一层中的分子将是具有基本功能的候选者，因此在微观水平上分析石头基质是必要的。为此，将肾结石脱矿、切片、染色并通过显微镜成像，使用微型CT进行精确定位。组织学染色显示结石内相邻层密度的异质性。其他结果也显示了辉煌的和独特的自发荧光模式在脱钙肾石，表明在相邻层的有机成分不均匀。使用激光显微切割（LMD）切割草酸钙（CaOx）结石区域进行蛋白质分析。广泛区域的脱矿钙氧化石部分的LMD产生相同的蛋白质中发现的不同标本的粉碎钙氧化石。这些创新的方法将允许在异质性结石基质内的蛋白质组成的空间映射。在空间上与矿物质沉积一致的蛋白质将成为结石生长所必需的分子的候选者。这种分析将需要评估结石基质中的数千种蛋白质中的哪一种可能是结石生长的基础。我们的创新方法描述了一种新的实验室方法来研究人类肾结石，通过显微镜分析结石基质来揭示其分子空间异质性。我们的目的是，本手稿中提供的数据将促进未来对结石基质的分子质询，以便更好地了解导致肾结石形成的潜在机制。
The molecular mechanisms by which kidney stones grow are largely unknown. Organic molecules from the urine combine with mineral crystals to form stones, but analysis of the stone matrix has revealed over a thousand different proteins, with no clues as to which are important for stone growth. Molecules that are present in every layer of a stone would be candidates for having an essential function, and thus the analysis of the stone matrix at a microscopic level is necessary. For this purpose, kidney stones were demineralized, sectioned, stained, and imaged by microscopy, using micro CT for precise orientation. Histological staining demonstrated heterogeneity in the density of adjacent layers within stones. Additional results also showed brilliant and unique autofluorescence patterns in decalcified nephroliths, indicating heterogeneous organic composition in adjacent layers. Regions of calcium oxalate (CaOx) stones were dissected using laser microdissection (LMD) for protein analysis. LMD of broad regions of demineralized CaOx stone sections yielded the same proteins as those found in different specimens of pulverized CaOx stones. These innovative methodologies will allow spatial mapping of protein composition within the heterogeneous stone matrix. Proteins that consistently coincide spatially with mineral deposition would be candidates for molecules essential for stone growth. This kind of analysis will be required to assess which of the thousand proteins in the stone matrix may be fundamental for stone growth. Our innovative methodology describes a novel laboratory method to study human kidney stones, whereby the stone matrix is microscopically analyzed to reveal its molecular spatial heterogeneity. It is our intent that the data presented in this manuscript will foster future molecular interrogations into the stone matrix for a better understanding of the underlying mechanisms contributing to renal stone formation.
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影响因子：
4.5
作者：
Ananth, K;Kavanagh, JP;Rao, PN
通讯作者：
Rao, PN
影响因子：
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作者：
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通讯作者：
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通讯作者：
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