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Updated: Jun 3, 2026
04:34
Development and Evaluation of a Rat Model of Full-Thickness Cartilage Defects
Published on: May 19, 2023
Modified house-keeping gene expression in a rat tail compression loading-induced disc degeneration model
Takashi Yurube1, Toru Takada, Hiroaki Hirata
1Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan.
Summary
House-keeping genes (HKGs) are vital for normalizing gene expression. This study found Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) suitable for intervertebral disc research, while β-actin is not recommended due to loading-induced changes.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Regenerative Medicine
Background:
- House-keeping genes (HKGs) are essential for normalizing gene expression in molecular studies.
- The suitability of commonly used HKGs in intervertebral disc degeneration research remains controversial.
Purpose of the Study:
- To evaluate the reliability of five common HKGs for gene quantification in nucleus pulposus cells using a rat tail static compression model.
- To identify a suitable HKG for gene expression analysis in intervertebral disc degeneration research.
Main Methods:
- A prevalidated rat tail static compression model was used to induce disc degeneration.
- Real-time RT-PCR and immunofluorescence were employed to analyze the expression of five HKGs (β-actin, β-glucuronidase, β-2 microglobulin, GAPDH, and LDHA) under static compression.
- Messenger RNA (mRNA) and protein levels of HKGs were assessed over 56 days.
Main Results:
- β-2 microglobulin and GAPDH showed consistent mRNA expression levels.
- β-glucuronidase was slightly up-regulated, while β-actin and lactate dehydrogenase A (LDHA) were significantly down-regulated.
- β-actin mRNA and protein levels drastically decreased under compression, becoming undetectable by day 28.
- GAPDH maintained stable mRNA and protein expression throughout the experiment.
Conclusions:
- β-actin and potentially LDHA are not appropriate as internal references in disc cells under prolonged static compression.
- GAPDH is a feasible and reliable HKG for internal normalization in gene quantification studies of intervertebral disc cells.
- These findings provide crucial guidance for selecting appropriate endogenous controls in disc degeneration research.
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