Maintaining mRNA integrity during decalcification of mineralized tissues.
Maintaining mRNA integrity during decalcification of mineralized tissues.
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DOI：
10.1371/journal.pone.0058154
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Bateman JF
中科院分区：
文献类型：
--
作者：
Belluoccio D;Rowley L;Little CB;Bateman JF
细胞外基质的生物矿化在许多病理状况（例如癌症和血管疾病）中不适当地发生，但在正常哺乳动物发育期间，钙化仅限于骨骼和牙列的形成。传统的脱钙/固定方法会导致 mRNA 显着降解，从而影响了对矿化骨骼组织中基因表达的综合研究。在这项研究中，我们开发了一种新型 RNAlater/EDTA 脱钙方法，可以保护成熟小鼠胫骨骨骺中 mRNA 的完整性。此外，除了免疫组织学和原位组织学之外，该方法还保留了组织结构，以便通过组织学切片和显微解剖来确定区域特异性基因表达。该方法将广泛适用于各种病理条件下钙化组织的分子分析，并且对于解剖发育过程中的小鼠骨和关节组织以及关节炎等重要临床条件下的基因表达特别重要。
Biomineralization of the extracellular matrix occurs inappropriately in numerous pathological conditions such as cancer and vascular disease, but during normal mammalian development calcification is restricted to the formation of the skeleton and dentition. The comprehensive study of gene expression in mineralized skeletal tissues has been compromized by the traditional decalcification/fixation methods that result in significant mRNA degradation. In this study we developed a novel RNAlater/EDTA decalcification method that protects the integrity of the mRNA in mature mouse tibial epiphyses. Furthermore, this method preserves the tissue structure to allow histological sectioning and microdissection to determine region-specific gene expression, in addition to immuno- and in situ histology. This method will be widely applicable to the molecular analysis of calcified tissues in various pathological conditions, and will be of particular importance in dissection of the gene expression in mouse bone and joint tissues during development and in important clinical conditions such as arthritis.
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影响因子：
3.7
作者：
Cameron TL;Bell KM;Tatarczuch L;Mackie EJ;Rajpar MH;McDermott BT;Boot-Handford RP;Bateman JF
通讯作者：
Bateman JF
影响因子：
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通讯作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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