Oguz Erbil, Tsai Tsung-Ting, Di Martino Alberto, Guttapalli Asha, Albert Todd J, Shapiro Irving M, Risbud Makarand V
Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Spine (Phila Pa 1976). 2007 Jan 1;32(1):9-16. doi: 10.1097/01.brs.0000250302.74574.98.
Galectin-3 expression in rat intervertebral disc at different stages in postnatal life is evaluated.
To determine if galectin-3 expression is confined to cells of the nucleus pulposus in the postnatal rat intervertebral disc.
During embryonic development, the anulus fibrosus is derived from the sclerotome, whereas the nucleus pulposus is notochordal. Many authorities opine that in the postnatal disc, notochordal cells play a central role in controlling the development of degenerative disc disease. Surprisingly, unequivocal evidence supporting the existence of notochordal cells in the nucleus pulposus in postnatal life has yet to be demonstrated. Since the expression of galectin-3 is commonly used to identify notochordal cells, we evaluated its expression in tissues of the rat disc and in cultured cells.
Galectin-3 expression was studied in the nucleus pulposus and anulus fibrosus tissue of rat discs (2 days, 9 weeks, and 10 months old), and cultured cells using different biochemical and molecular biology methods. Rat sternal cartilage and cultured sternal chondrocytes were used as controls.
Immunohistochemical studies indicated that galectin-3 was present in the nucleus pulposus and anulus fibrosus. In both discal tissues and cultured cells, studies confirmed that there was a robust expression of galectin-3 messenger ribonucleic acid and protein. Protein expression patterns were similar in neonatal, young, and mature rats. There was also evidence of intracellular and membrane expression of galectin-3 in the cultured disc cells. Finally, significant levels of galectin-3 were evident in rat sternal cartilage and cultured sternal chondrocytes.
Results of the study indicate that galectin-3 is expressed in the neonatal, young, and mature rat disc, and its expression is not restricted to the cells of the nucleus pulposus. Because of its ubiquitous expression, this protein cannot be used as a marker of notochordal cells in the postnatal rat disc.
评估出生后不同阶段大鼠椎间盘内半乳糖凝集素-3的表达情况。
确定半乳糖凝集素-3的表达是否局限于出生后大鼠椎间盘的髓核细胞。
在胚胎发育过程中,纤维环源自硬骨节,而髓核源自脊索。许多权威人士认为,在出生后的椎间盘里,脊索细胞在控制椎间盘退变疾病的发展中起核心作用。令人惊讶的是,支持出生后生活中髓核内存在脊索细胞的明确证据尚未得到证实。由于半乳糖凝集素-3的表达通常用于识别脊索细胞,我们评估了其在大鼠椎间盘组织和培养细胞中的表达。
采用不同的生化和分子生物学方法,研究大鼠椎间盘(2日龄、9周龄和10月龄)的髓核和纤维环组织以及培养细胞中的半乳糖凝集素-3表达。以大鼠胸骨软骨和培养的胸骨软骨细胞作为对照。
免疫组织化学研究表明,半乳糖凝集素-3存在于髓核和纤维环中。在椎间盘组织和培养细胞中,研究证实半乳糖凝集素-3信使核糖核酸和蛋白质有强烈表达。新生、幼年和成年大鼠的蛋白质表达模式相似。在培养的椎间盘细胞中也有半乳糖凝集素-3细胞内和膜表达的证据。最后,在大鼠胸骨软骨和培养的胸骨软骨细胞中明显存在高水平的半乳糖凝集素-3。
研究结果表明,半乳糖凝集素-3在新生、幼年和成年大鼠椎间盘中均有表达,且其表达不限于髓核细胞。由于其广泛表达,这种蛋白质不能用作出生后大鼠椎间盘中脊索细胞的标志物。