Article

Microarray analysis of perichondral and reserve growth plate zones identifies differential gene expressions and signal pathways

Musculoskeletal Sciences Research Laboratory, Department of Orthopedic Surgery, State University of New York Upstate Medical University, Syracuse, New York, 13210, USA.
Bone (Impact Factor: 4.46). 06/2008; 43(3):511-20. DOI: 10.1016/j.bone.2008.04.021
Source: PubMed

ABSTRACT In the growth plate, the reserve and perichondral zones have been hypothesized to have similar functions, but their exact functions are poorly understood. Our hypothesis was that significant differential gene expression exists between perichondral and reserve chondrocytes that may differentiate the respective functions of these two zones. Normal Sprague-Dawley rat growth plate chondrocytes from the perichondral zone (PC) and reserve zone (RZ) were isolated by laser microdissection and then subjected to microarray analysis. In order to most comprehensively capture the unique features of the two zones, we analyzed both the most highly expressed genes and those that were most significantly different from the proliferative zone (PZ) as a single comparator. Confirmation of the differential expression of selected genes was done by quantitative real-time RT-PCR. A total of 8 transcripts showing high expression unique to the PC included translationally-controlled tumor protein (Tpt1), connective tissue growth factor (Ctgf), mortality factor 4 (Morf4l1), growth arrest specific 6 (Gas6), type V procollagen (Col5a2), frizzled-related protein (Frzb), GDP-dissociation inhibitor 2 (Gdi2) and Jun D proto-oncogene (Jund). In contrast, 8 transcripts showing unique high expression in the RZ included hyaluronan and proteoglycan link protein 1 (Hapln1), hemoglobin beta-2 subunit, type I procollagen (Col1a2), retinoblastoma binding protein 4 (LOC685491), Sparc-related modular calcium binding 2 (Smoc2), and calpastatin (Cast). Other genes were highly expressed in cells from both PC and RZ zones, including collagen II, collagen IX, catenin (cadherin associated protein) beta 1, eukaryotic translation elongation factor, high mobility group, ribosomal protein, microtubule-associated protein, reticulocalbin, thrombospondin, retinoblastoma binding protein, carboxypeptidase E, carnitine palmitoyltransferase 1, cysteine rich glycoprotein, plexin B2 (Plxnb2), and gap junction membrane channel protein. Functional classification of the most highly expressed transcripts were analyzed, and the pathway analysis indicated that ossification, bone remodeling, and cartilage development were uniquely enriched in the PC whereas both the PC and RZ showed pathway enrichment for skeletal development, extracellular matrix structural constituent, proteinaceous extracellular matrix, collagen, extracellular matrix, and extracellular matrix part pathways. We conclude that differential gene expression exists between the RZ and PC chondrocytes and these differentially expressed genes have unique roles to play corresponding to the function of their respective zones.

Download full-text

Full-text

Available from: Frank Middleton, Sep 02, 2015
0 Followers
 · 
90 Views
 · 
36 Downloads
  • Source
    • "Interestingly, previous studies have suggested a role for CPE in skeletal development. These studies demonstrated abundant expression of CPE in developing skeletal structures (Zheng et al., 1994) and growth plate chondrocytes (Zhang et al., 2008), although its specific role was not determined in these studies. Recently, Cawley et al. (2010) reported that CPE indirectly regulates bone metabolism through the sympathetic nervous system. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Osteoclasts are large polykaryons that have the unique capacity to degrade bone and are generated by the differentiation of myeloid lineage progenitors. To identify the genes involved in osteoclast development, we performed microarray analysis, and we found that carboxypeptidase E (CPE), a prohormone processing enzyme, was highly upregulated in osteoclasts compared with their precursors, bone marrow-derived macrophages (BMMs). Here, we demonstrate a novel role for CPE in receptor activator of NF-kaapB ligand (RANKL)-induced osteoclast differentiation. The overexpression of CPE in BMMs increases the formation of tartrate- resistant acid phosphatase (TRAP)-positive multinuclear osteoclasts and the expression of c-Fos and nuclear factor of activated T cells c1 (NFATc1), which are key regulators in osteoclastogenesis. Furthermore, employing CPE knockout mice, we show that CPE deficiency attenuates osteoclast formation. Together, our data suggest that CPE might be an important modulator of RANKL-induced osteoclast differentiation.
    Molecules and Cells 09/2014; 37(9). DOI:10.14348/molcells.2014.0179 · 2.24 Impact Factor
  • Source
    • "While gene expression profiling is not generally reported for the epiphyseal growth plate mediated calcification, there are reports for gene expression in the early differentiation of mesenchymal cells into chondrocytes [Cameron et al., 2009], in the reserve [Zhang et al., 2008], proliferating and hypertrophic zones of the growth plate [Wang et al., 2004b], and in healing fracture callus development [Rundle et al., 2006; Wang et al., 2006; Khan et al., 2008] a process that mimics endochondral ossification. Similarly there are reports of gene expression during intramembranous bone formation [Kuroda et al., 2005], and for the effects of different factors on growth plate chondrocytes [Ulici et al., 2010]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Chick limb-bud mesenchymal stem cells plated in high density culture in the presence of 4 mM inorganic phosphate and vitamin C differentiate and form a mineralizable matrix, resembling that of the chick growth plate. To further elucidate the mechanism that allows these cultures to form physiologic hydroxyapatite deposits, and how the process can be manipulated to gain insight into mineralization mechanisms, we compared gene expression in mineralizing (with 4 mM inorganic phosphate) and non-mineralizing cultures (containing only 1 mM inorganic phosphate) at the start of mineralization (day 11) and after mineralization reached a plateau (day 17) using a chick specific microarray. Based on replicate microarray experiments and K-cluster analysis, several genes associated with the mineralization process were identified, and their expression patterns confirmed throughout the culture period by quantitative RT-PCR. The functions of bone morphogenetic protein 1, BMP1, dentin matrix protein 1, DMP1, the sodium phosphate co-transporter, NaPi IIb, matrix metalloprotease 13. MMP-13, and alkaline phosphatase, along with matrix protein genes (type X collagen, bone sialoprotein, and osteopontin) usually associated with initiation of mineralization are discussed.
    Journal of Cellular Biochemistry 02/2011; 112(2):607-13. DOI:10.1002/jcb.22951 · 3.37 Impact Factor
  • Advances in Psychology, pages 431-451;
Show more