Identification of differentially expressed genes between osteoblasts and osteocytes.
Identification of differentially expressed genes between osteoblasts and osteocytes.
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DOI：
10.1016/j.bone.2009.06.010
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发表时间：
2009-10
期刊：
影响因子：
4.1
通讯作者：
Kalajzic, Ivo
中科院分区：
文献类型：
--
作者：
Paic, Frane;Igwe, John C.;Nori, Ravi;Kronenberg, Mark S.;Franceschetti, Tiziana;Harrington, Patrick;Kuo, Lynn;Shin, Dong-Guk;Rowe, David W.;Harris, Stephen E.;Kalajzic, Ivo
骨细胞是哺乳动物骨骼中含量最丰富的细胞成分，在骨量维持和重建中具有重要作用。为了阐明成骨细胞和骨细胞之间的差异基因表达，我们对它们的基因图谱进行了全面的分析。这两个成熟群体的选择性鉴定是利用骨系细胞的视觉标记物实现的。我们利用了双GFP报告小鼠，其中骨细胞表达由DMP1启动子引导的GFP(黄玉)，而成骨细胞通过由2.3kb的Col1a1启动子驱动的GFP(青色)的表达来鉴定。7日龄新生颅骨组织学分析证实DMP1GFP在骨细胞中的表达模式，Col2.3在成骨细胞和骨细胞中的表达模式。为了分离不同的细胞群，我们使用了荧光激活细胞分选(FACS)。细胞悬液提取RNA，体外转录和标记基因，用Illumina WG-6v1微珠芯片分析基因表达。在对四个生物重复的原始数据进行归一化后，发现3444个基因在所有三个分类的细胞群中存在：GFP阴性、Col2.3cyan+(成骨细胞)和DMP1topaz+(前骨细胞和骨细胞)。我们介绍了DMP1topaz+和Col2.3 Cyan+细胞之间基因表达变化超过2倍的基因。根据相关基因本体论术语对所选基因进行分类和分组。成骨和骨骼发育相关基因Bmp4、Bmp8a、Dmp1、Enpp1、PHEX和Ank在DMP1topaz+细胞中高表达。在DMP1topaz+细胞中，大多数编码细胞外基质成分和分泌蛋白的基因表达降低，而大多数编码质膜蛋白的基因表达增加。有趣的是，在DMP1topaz+细胞中，大量与肌肉发育和功能以及与神经元表型相关的基因增加，这表明骨细胞生物学的一些新方面。虽然本研究中大量在DMP1topaz+和Col2.3cyan+细胞中差异表达的基因已经被归类到骨发育和生理中，但对于它们中的大多数我们仍然缺乏实质性的数据。因此，分离骨细胞和成骨细胞群体及其随后的微阵列分析使我们能够确定一些或多个基因和途径，这些基因和途径可能在骨量调节中发挥作用。
Osteocytes represent the most abundant cellular component of mammalian bones with important functions in bone mass maintenance and remodeling. To elucidate the differential gene expression between osteoblasts and osteocytes we completed a comprehensive analysis of their gene profiles. Selective identification of these two mature populations was achieved by utilization of visual markers of bone lineage cells. We have utilized dual GFP reporter mice in which osteocytes are expressing GFP (topaz) directed by the DMP1 promoter, while osteoblasts are identified by expression of GFP (cyan) driven by 2.3kb of the Col1a1 promoter. Histological analysis of 7-day-old neonatal calvaria confirmed the expression pattern of DMP1GFP in osteocytes and Col2.3 in osteoblasts and osteocytes. To isolate distinct populations of cells we utilized fluorescent activated cell sorting (FACS). Cells suspensions were subjected to RNA extraction, in vitro transcription and labeling of cDNA and gene expression was analyzed using the Illumina WG-6v1 BeadChip. Following normalization of raw data from four biological replicates, 3444 genes were called present in all three sorted cell populations: GFP negative, Col2.3cyan+ (osteoblasts), and DMP1topaz+(preosteocytes and osteocytes). We present the genes that showed in excess of a 2-fold change for gene expression between DMP1topaz+ and Col2.3cyan+ cells. The selected genes were classified and grouped according to their associated gene ontology terms. Genes clustered to osteogenesis and skeletal development such as Bmp4, Bmp8a, Dmp1, Enpp1, Phex and Ank were highly expressed in DMP1topaz+cells. Most of the genes encoding extracellular matrix components and secreted proteins had lower expression in DMP1topaz+ cells, while most of the genes encoding plasma membrane proteins were increased. Interestingly a large number of genes associated with muscle development and function and with neuronal phenotype were increased in DMP1topaz+ cells, indicating some new aspects of osteocyte biology. Although a large number of genes differentially expressed in DMP1topaz+ and Col2.3cyan+ cells in our study have already been assigned to bone development and physiology, for most of them we still lack any substantial data. Therefore, isolation of osteocyte and osteoblast cell populations and their subsequent microarray analysis allowed us to identify a number or genes and pathways with potential roles in regulation of bone mass.
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DOI：
10.1152/ajpendo.00592.2005
发表时间：
2006-07-01
影响因子：
5.1
作者：
Govoni, Kristen E.;Lee, Seong Keun;Mohan, Subburaman
通讯作者：
Mohan, Subburaman
影响因子：
--
作者：
Holmberg, Johan;Ingner, Gorel;Johansson, Curt;Leander, Peter;Hjalt, Tord A.
通讯作者：
Hjalt, Tord A.
影响因子：
4.8
作者：
Luo, Q;Kang, Q;He, TC
通讯作者：
He, TC
影响因子：
7.6
作者：
Feng, JQ;Huang, H;Mishina, Y
通讯作者：
Mishina, Y
影响因子：
4.8
作者：
Hassan, Mohammad Q.;Tare, Rahul S.;Lian, Jane B.
通讯作者：
Lian, Jane B.