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| Status |
Public on Mar 24, 2009 |
| Title |
Asporin is Expressed at Higher Levels in the More Degenerate Human Intervertebral Disc Tissue |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Asporin, also known as periodontal ligament-associated protein 1 (PLAP1), is a member of the family of small leucine-rich proteoglycan (SLRP) family. It is present within the cartilage extracellular matrix (ECM), and is reported have a genetic association with osteoarthritis. Its D14 allele has recently been found to be associated with lumbar disc degeneration in Asian subjects. There have been no studies, however, of this gene’s normal immunohistochemical localization within the human intervertebral disc, nor of expression levels in Caucasian individuals with disc degeneration. Studies were approved by our human subjects Institutional Review Board. Methods included immunohistochemical localization of asporin in the disc of humans and the sand rat (a small rodent with spontaneous age-related disc degeneration), and Affymetrix microarray analysis of asporin gene expression in vivo and in vitro.
mmunohistochemical studies of human discs revealed that some, but not all, cells of the outer annulus expressed asporin. Fewer cells in the inner annulus contained asporin, and it was rarely present in cells in the nucleus pulposus. Similar patterns were found for the presence of asporin in lumbar discs of sand rats. Substantial relative gene expression levels were seen for asporin in both disc tissue and in annulus cells grown in three-dimensional culture. More degenerate human discs (Thompson grade 4) showed higher expression levels of asporin than did less degenerate (grade 1, 2 and 3) discs, p = 0.004. In the discs of Caucasian subjects studied here, and in the sand rat, greater immunolocalization levels were found in the outer compared to inner annulus. Localization was rare in the nucleus. Gene expression studies showed greatest expression of asporin in the more degenerate human discs in vivo.
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| Overall design |
Disc Tissue samples were obtained via the National Cancer Institute Cooperative Tissue Network (CHTN) as well as surgical disc procedures performed on patients with herniated discs and degenerative disc disease. Tissue was fixed and paraffin embedded. Standard laser capture microdissection (LCM) techniques were used to collect the cells from which total RNA was isolated and analyzed via microarray. Based on the Thompson scouring system, unhealthy discs (grade 4) were compared to healthy discs (grades 2,3).
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| Contributor(s) |
Gruber HE, Ingram JA, Hoelscher GL, Zinchenko N, Hanley EN Jr, Sun Y |
| Citation(s) |
19535298 |
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| Submission date |
Mar 13, 2009 |
| Last update date |
Mar 21, 2012 |
| Contact name |
Helen Gruber |
| E-mail(s) |
Helen.Gruber@carolinashealthcare.org
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| Organization name |
Carolinas HealthCare System
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| Department |
Orthopaedic Surgery
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| Lab |
Orthopaedic Research
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| Street address |
1542 Graden Terrace
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| City |
Charlotte |
| State/province |
NC |
| ZIP/Postal code |
28203 |
| Country |
USA |
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| Platforms (1) |
| GPL1352 |
[U133_X3P] Affymetrix Human X3P Array |
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| Samples (15)
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| Relations |
| BioProject |
PRJNA116483 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE15227_RAW.tar |
101.0 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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