Osmolarity regulates gene expression in intervertebral disc cells determined by gene array and real-time quantitative RT-PCR.
Intervertebral disc (IVD) cells experience a broad range of physicochemical stimuli under physiologic conditions, including alterations in their osmotic environment. Cellular responses to altered osmolarity have been documented at the transcriptional and post-translational level, but mainly for extracellular matrix proteins. In this study, the gene expression profile of human IVD cells was quantified with gene array technology following exposure to increased osmolarity in order to capture the biological responses for a broad set of targets. A total of 42 genes were identified in IVD cells as significantly changed following culture under hyper-osmotic conditions. Gene expression patterns were verified using RT-PCR. Genes identified in this study include those related to cytoskeleton remodeling and stabilization (ephrin-B2, muskelin), as well as membrane transport (ion transporter SLC21A12, osmolyte transporter SLC5A3, monocarboxylic acid SLC16A6). An unexpected finding was the differential regulation of the gene for the neurotrophin, brain-derived neurotrophic factor, by hyper-osmotic stimuli that suggests a capability of IVD cells to respond to physicochemical stimuli with factors that may regulate discogenic pain.
Boyd
Lawrence M
LM
Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708, USA.
Richardson
William J
WJ
Chen
Jun
J
Kraus
Virginia B
VB
Tewari
Alok
A
Setton
Lori A
LA
D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology / Q000502:physiology*; D008297:Male; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D009994:Osmolar Concentration; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods