Spatial and Temporal Regulation of Gene Expression in the Mammalian Growth Plate
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ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE16981 | GEO | 2010/01/28
SECONDARY ACCESSION(S): PRJNA117283
REPOSITORIES: GEO
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Project description:To explore the mechanisms responsible for spatial and temporal regulation of the growth plate, we microdissected postnatal rat growth plates into their constituent zones and then used microarray analysis to characterize the changes in gene expression that occur as chondrocytes undergo spatially-associated differentiation and temporally-associated senescence. Microdissection was used to collect individual growth plate zones from proximal tibiae of 1-wk rats and the proliferative and early hypertrophic zones of growth plates from 3-, 6-, 9-, and 12-wk rats and gene expression was analyzed using microarray.
2010-02-22 | E-GEOD-16981 | biostudies-arrayexpress
Project description:Axial growth of long bones occurs through a coordinated process of growth plate chondrocyte proliferation and differentiation. This maturation of chondrocytes is reflected in a zonal change in gene expression and cell morphology from resting to proliferative, prehypertrophic, and hypertrophic chondrocytes of the growth plate followed by ossification. A major experimental limitation in understanding growth plate biology and pathophysiology is the lack of a robust technique to isolate cells from the different zones, particularly from small animals. Here, we report on a new strategy for separating distinct chondrocyte populations from mouse growth plates. By transcriptome profiling of microdissected zones of growth plates, we identified novel, zone-specific cell surface markers and used these for flow cytometry and immunomagnetic cell separation to quantify, enrich, and characterize chondrocytes populations with respect to their differentiation status. This approach provides a novel platform to study cartilage development and characterize mouse growth plate chondrocytes to reveal unique cellular phenotypes of the distinct subpopulations within the growth plate.
2020-02-29 | GSE144362 | GEO
Project description:In order to characterize mRNA expression in the growth plate, we microdissected postnatal rat growth plates into their constituent zones and used microarray analysis to assess the abundences of individual transcripts. Expression patterns of PTHrP and Ihh-related genes were confirmed using real-time PCR. Using a gli1-lacZ mouse, Gli1 expression, presumably representing Ihh signaling, was visualized during pre- and postnatal development.
2010-12-01 | GSE23432 | GEO
Project description:In order to characterize mRNA expression in the growth plate, we microdissected postnatal rat growth plates into their constituent zones and used microarray analysis to assess the abundences of individual transcripts. Expression patterns of PTHrP and Ihh-related genes were confirmed using real-time PCR. Using a gli1-lacZ mouse, Gli1 expression, presumably representing Ihh signaling, was visualized during pre- and postnatal development. Microdissection was used to collect individual growth plate zones from proximal tibiae of 1-wk rats and gene expression was analyzed using microarray.
2010-12-01 | E-GEOD-23432 | biostudies-arrayexpress
Project description:Chondrocytes advance through the long bone growth plate in synchrony, allowing their segregation into one of four zones each corresponding to a different phase of maturation. Beginning from the epiphysis the recognized zones are: reserve, proliferative, prehypertrophic, and hypertrophic. A fifth zone, the perichondrium is located in apposition to the growth plate at its circumferential surface. The progression of chondrocytes through these zones, and hence growth, is regulated by a complex interplay of signaling pathways, defects in which lead to osteochondrodysplasias. At present, there are over 370 recognized skeletal disorders, slightly more than half of which have been explained at the molecular level. Through a combination of laser microdissection, linear mRNA amplification, and GeneChip analysis, our laboratory has acquired the zone specific expression profile of chondrocytes from the distal femoral growth plates of two normal patients. On average, transcripts interrogated by 12,193 or 18,454 GeneChip probe sets (out of a possible 54,000 probe sets) were present in patients 1 and 2, respectively. The homologous inter-array correlations for the log(2) intensities were ≥ 0.91±0.01. Keywords: Chondrocyte differentiation program.
2013-09-13 | GSE9160 | GEO
Project description:Articular and growth plate cartilage have comparable structures consisting of three distinct layers of chondrocytes, suggesting similar differentiation programs and therefore similar gene expression profiles. To address this hypothesis and to explore transcriptional changes that occur during the onset of articular and growth plate cartilage divergence, we used microdissection of 10-day-old rat proximal tibial epiphyses, microarray analysis, and bioinformatics to compare gene expression profiles in individual layers of articular and growth plate cartilage. We found that many genes that were spatially upregulated in intermediate/deep zone of articular cartilage were also spatially upregulated in resting zone of growth plate cartilage (overlap greater than expected by chance, P < 0.001). Interestingly, superficial zone of articular cartilage showed an expression profile with similarities to both proliferative and hypertrophic zones of growth plate cartilage (P < 0.001 each). Additionally, significant numbers of known proliferative zone markers (3 out of 6) and hypertrophic zone markers (27 out of 126) were spatially upregulated in superficial zone compared to intermediate/deep zone (more than expected by chance, P < 0.001 each). In conclusion, we provide evidence that intermediate/deep zone of articular cartilage has a gene expression profile more similar to resting zone of growth plate cartilage, whereas superficial zone has a gene expression profile more similar to proliferative and hypertrophic zones. 10-day-old rat proximal tibial epiphyses were manually microdissected into articular cartilage superficial (SZ) and intermediate/deep (IDZ) zones and growth plate cartilage resting zone (RZ) for total RNA extraction and hybridization on Affymetrix microarrays. We used 10-day-old animals because, at this age, the secondary ossification center has recently begun to form and divides the epiphysis into articular cartilage distally and growth plate cartilage more centrally. The 4 SZ samples were taken from animals 5-8, respectively, whereas the 4 IDZ and 4 RZ samples were each taken from animals 1-2, 3-4, 5-6, and 7-8, respectively.
2014-08-16 | E-GEOD-54216 | biostudies-arrayexpress
Project description:Chondrocytes advance through the long bone growth plate in synchrony, allowing their segregation into one of four zones each corresponding to a different phase of maturation. Beginning from the epiphysis the recognized zones are: reserve, proliferative, prehypertrophic, and hypertrophic. A fifth zone, the perichondrium is located in apposition to the growth plate at its circumferential surface. The progression of chondrocytes through these zones, and hence growth, is regulated by a complex interplay of signaling pathways, defects in which lead to osteochondrodysplasias. At present, there are over 370 recognized skeletal disorders, slightly more than half of which have been explained at the molecular level. Through a combination of laser microdissection, linear mRNA amplification, and GeneChip analysis, our laboratory has acquired the zone specific expression profile of chondrocytes from the distal femoral growth plates of two normal patients. On average, transcripts interrogated by 12,193 or 18,454 GeneChip probe sets (out of a possible 54,000 probe sets) were present in patients 1 and 2, respectively. The homologous inter-array correlations for the log(2) intensities were M-bM-^IM-% 0.91M-BM-10.01. Keywords: Chondrocyte differentiation program. Populations of cells corresponding to the hypertrophic, prehypertrophic, proliferative, reserve, and perichondrium, were separately isolated by laser microdissection from cryostat sections of human growth plates using a Leica AS-LMD microscope. RNA was prepared from the isolated cells, amplified using a T7 linear amplification protocol, and analyzed by hybridization to Affymetrix microarrays. The distal femoral growth plate (left or right) was exclusively used for this study. Patient 1 of this study is an 11-10/12 year old Caucasian female and patient 2 is a 13-3/12 year old Caucasian male.
2013-09-13 | E-GEOD-9160 | biostudies-arrayexpress
Project description:Articular and growth plate cartilage have comparable structures consisting of three distinct layers of chondrocytes, suggesting similar differentiation programs and therefore similar gene expression profiles. To address this hypothesis and to explore transcriptional changes that occur during the onset of articular and growth plate cartilage divergence, we used microdissection of 10-day-old rat proximal tibial epiphyses, microarray analysis, and bioinformatics to compare gene expression profiles in individual layers of articular and growth plate cartilage. We found that many genes that were spatially upregulated in intermediate/deep zone of articular cartilage were also spatially upregulated in resting zone of growth plate cartilage (overlap greater than expected by chance, P < 0.001). Interestingly, superficial zone of articular cartilage showed an expression profile with similarities to both proliferative and hypertrophic zones of growth plate cartilage (P < 0.001 each). Additionally, significant numbers of known proliferative zone markers (3 out of 6) and hypertrophic zone markers (27 out of 126) were spatially upregulated in superficial zone compared to intermediate/deep zone (more than expected by chance, P < 0.001 each). In conclusion, we provide evidence that intermediate/deep zone of articular cartilage has a gene expression profile more similar to resting zone of growth plate cartilage, whereas superficial zone has a gene expression profile more similar to proliferative and hypertrophic zones.
2014-08-16 | GSE54216 | GEO
Project description:Proliferative zone chondrocytes were microdissected from control and Ift88-deleted growth plates to determine gene expression profiles regulated by primary cilia.
2013-01-02 | GSE35003 | GEO
Project description:Growth plate chondrocytes were isolated from the distal metacarpus of young dairy cattle (all under 10 mo of age), the chondrocytes were released from the extracellular matrix by digestion with Collagenase P for 4 hours, and the various zones of the growth plate were separated by density centrifugation. The least-dense Hypertrophic Zone (HZ) cells were compared to the most-dense Reserve Zone (RZ) cells. 6 pairs of HZ vs RZ were compared by microarray.
2009-10-27 | GSE18738 | GEO