A method for isolating high quality RNA from mouse cortical and cancellous bone.
A method for isolating high quality RNA from mouse cortical and cancellous bone.
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DOI：
10.1016/j.bone.2014.07.022
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发表时间：
2014-11
期刊：
影响因子：
4.1
通讯作者：
van der Meulen, Marjolein C. H.
中科院分区：
文献类型：
--
作者：
Kelly, Natalie H.;Schimenti, John C.;Ross, F. Patrick;van der Meulen, Marjolein C. H.
皮质骨-松质骨部位脆性骨折的高发病率，加上个体骨骼部位对负荷和疾病表现出不同的反应性这一事实，强调需要分别记录皮质骨和松质骨中的基因表达。另一个混杂因素是骨髓污染，因为其高细胞性可能影响基因表达测量。我们从完整小鼠胫骨的皮质骨和松质骨中分离出RNA，并在通过冲洗或离心去除骨髓后分离出RNA。通过每种方法从松质骨中分离的RNA足以用于基因表达分析。冲洗比冲洗更有效地去除污染细胞，如组织学所示，并降低了高表达的红细胞基因Icam 4的表达。相比之下，离心的皮质骨中骨相关基因Col 1a 1和Bglap的表达分别高出12倍和13倍，而无骨髓松质骨中的表达水平分别高出30倍和31倍。皮质骨中Col 1a 1和Bglap的表达高于松质骨。因此，通过这种新方法分离的RNA可以揭示皮质骨和松质骨部位基因表达的位点特异性变化。
The high incidence of fragility fractures in cortico-cancellous bone locations, plus the fact that individual skeletal sites exhibit different responsiveness to load and disease, emphasizes the need to document separately gene expression in cortical and cancellous bone. A further confounding factor is marrow contamination since its high cellularity may effect gene expression measurements. We isolated RNA from cortical and cancellous bone of intact mouse tibiae, and also after marrow removal by flushing or centrifugation. RNA isolated from cancellous bone by each method was sufficient for gene expression analysis. Centrifugation removed contaminating cells more efficiently than flushing, as indexed by histology and decreased expression of Icam4, a highly expressed erythroid gene. In contrast, centrifuged cortical bone had 12-and 13-fold higher expression of the bone-related genes Col1a1 and Bglap, while levels in marrow-free cancellous bone were 30-and 31-fold higher when compared to bone where marrow was left intact. Furthermore, cortical bone had higher expression of Col1a1 and Bglap than cancellous bone. Thus, RNA isolated by this novel approach can reveal site-specific changes in gene expression in cortical and cancellous bone sites.
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影响因子：
4.1
作者：
Lynch, Maureen E.;Main, Russell P.;Xu, Qian;Schmicker, Thomas L.;Schaffler, Mitchell B.;Wright, Timothy M.;van der Meulen, Marjolein C. H.
通讯作者：
van der Meulen, Marjolein C. H.
影响因子：
3.3
作者：
Lynch, Maureen E.;Main, Russell P.;van der Meulen, Marjolein C. H.
通讯作者：
van der Meulen, Marjolein C. H.
影响因子：
4.1
作者：
Wasserman, Elad;Webster, Duncan;Bab, Itai
通讯作者：
Bab, Itai
影响因子：
4.8
作者：
Armstrong, Victoria J.;Muzylak, Mariusz;Lanyon, Lance E.
通讯作者：
Lanyon, Lance E.
影响因子：
4.2
作者：
Holguin, Nilsson;Brodt, Michael D.;Sanchez, Michelle E.;Kotiya, Akhilesh A.;Silva, Matthew J.
通讯作者：
Silva, Matthew J.