title: Expression of semaphorin 3A and its receptors in the human intervertebral disc: potential role in regulating neural ingrowth in the degenerate intervertebral disc.
abstract: BACKGROUND
  Intervertebral disc (IVD) degeneration is considered a major underlying factor in the pathogenesis of chronic low back pain. Although the healthy IVD is both avascular and aneural, during degeneration there is ingrowth of nociceptive nerve fibres and blood vessels into proximal regions of the IVD, which may contribute to the pain. The mechanisms underlying neural ingrowth are, however, not fully understood. Semaphorin 3A (sema3A) is an axonal guidance molecule with the ability to repel nerves seeking their synaptic target. This study aimed to identify whether members of the Class 3 semaphorins were expressed by chondrocyte-like cells of the IVD addressing the hypothesis that they may play a role in repelling axons surrounding the healthy disc, thus maintaining its aneural condition.
  
  
  METHODS
  Human IVD samples were investigated using reverse transcription polymerase chain reaction (RT-PCR) to identify gene expression of sema3A, 3F and their receptors: neuropilins (1 and 2) and plexins (A1-4). Sema3A protein was also localised within sections of normal and degenerate human IVD and immunopositivity quantified. Serial sections were stained using PGP9.5 and CD31 to correlate semaphorin 3A expression with nerve and blood vessel ingrowth, respectively.
  
  
  RESULTS
  Sema3A protein was expressed highly in the healthy disc, primarily localised to the outer annulus fibrosus. In degenerate samples, sema3A expression decreased significantly in this region, although cell clusters within the degenerate nucleus pulposus exhibited strong immunopositivity. mRNA for sema3A receptors was also identified in healthy and degenerate tissues. CD31 and PGP9.5 were expressed most highly in degenerate tissues correlating with low expression of sema3A.
  
  
  CONCLUSIONS
  This study is the first to establish the expression of semaphorins and their receptors in the human IVD with a decrease seen in the degenerate painful IVD. Sema3A may therefore, amongst other roles, act as a barrier to neuronal ingrowth within the healthy disc.
authors.affiliation: Tissue Injury and Repair Group, School of Clinical and Laboratory Sciences, University of Manchester, Stopford Building, Oxford Road, Manchester, M13 9PT, UK. Sotonye.tolofari@student.manchester.ac.uk
authors.forename: Sotonye K
authors.initials: SK
authors.lastname: Tolofari
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Richardson
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Freemont
authors.forename: Judith A
authors.initials: JA
authors.lastname: Hoyland
mesh_terms: D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D007403:Intervertebral Disc / Q000294:innervation* / Q000378:metabolism* / Q000473:pathology; D055959:Intervertebral Disc Degeneration / Q000378:metabolism* / Q000473:pathology; D009414:Nerve Growth Factors / Q000096:biosynthesis; D039942:Neuropilin-1 / Q000096:biosynthesis*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D040121:Semaphorin-3A / Q000096:biosynthesis*; D043222:Ubiquitin Thiolesterase / Q000096:biosynthesis
chemical_list: D009414:Nerve Growth Factors; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D040121:Semaphorin-3A; C482860:UCHL1 protein, human; D039942:Neuropilin-1; D043222:Ubiquitin Thiolesterase