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首页> 外文期刊>Spine >Evidence of enhanced expression of osteopontin in spinal hyperostosis of the twy mouse.
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Evidence of enhanced expression of osteopontin in spinal hyperostosis of the twy mouse.

机译：骨桥蛋白在twy小鼠脊柱肥大中表达增强的证据。

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STUDY DESIGN: Gene expression and protein localization of osteopontin (OPN) in spinal hyperostosis of the twy mouse by means of in situ hybridization, immunohistochemistry, and Northern blot analysis. OBJECTIVE: To verify the involvement of OPN in spinal hyperostosis in the twy mouse and elucidate its ossification pattern at molecular levels. SUMMARY OF BACKGROUND DATA: OPN is a molecule that consistently colocalizes with ectopic calcification in human pathologic conditions. The twy mouse, which shows ectopic calcification of the spinal ligament resulting in hind limb paralysis, is considered to be a model for human ossification of the posterior longitudinal ligament of the spine. METHODS: Twenty-eight each of age-matched twy, heterozygote, and wild-type mice were killed at 2, 4, 8, 12, and 16 weeks old and subject to histologic and/or molecular analyses. Sections were hybridized with RNA probes for OPN and also stained with anti-OPN antibodies. Total cellular RNA was extracted from the cervicothoracic spine of each genotype at 2- and 16-week-old, and gene expression for OPN and COL10A1 was quantified by Northern blot analysis. RESULTS: Enhanced expression of OPN mRNA was observed in spinal hyperostotic lesions of the twy mouse, specifically in cells of the spinal ligament and chondrogenic cells in the outer layer of the anulus fibrosus. These trends were also confirmed by immunohistochemical analyses. Northern blot analysis showed that a considerable amount of OPN transcripts was detected in all genotypes at 2 weeks old, but the robust expression of OPN mRNA was maintained only in twy mice at 16 weeks old. COL10A1 transcripts were hardly detected regardless of the genotype at 16 weeks old. CONCLUSION: OPN was overexpressed in the hyperostotic spinal lesions of twy mice, and the hyperostosis was induced mainly by ectopic ossification of the spinal ligament. Because OPN is considered to be an inhibitor of calcification, further studies will be necessary to verify whether OPN overexpressed in the twy mouse is functional.
机译：研究设计：通过原位杂交，免疫组织化学和Northern印迹分析，骨质疏松蛋白（OPN）在twy小鼠脊柱肥大中的基因表达和蛋白定位。目的：验证OPN参与twy小鼠脊柱骨增生症并从分子水平阐明其骨化模式。背景数据摘要：OPN是在人类病理条件下始终与异位钙化共定位的分子。显示出导致后肢麻痹的脊髓韧带异位钙化的twy小鼠被认为是人的脊柱后纵韧带骨化的模型。方法：在28、2、4、8、12和16周龄时，分别杀死28只年龄匹配的twy，杂合子和野生型小鼠，并进行组织学和/或分子分析。将切片与OPN的RNA探针杂交，并用抗OPN抗体染色。在2周和16周龄时从每种基因型的颈胸椎提取总细胞RNA，并通过Northern印迹分析定量OPN和COL10A1的基因表达。结果：在twy小鼠的脊柱肥大性病变中，特别是在纤维环外层的脊膜韧带细胞和软骨细胞中，OPN mRNA的表达增强。免疫组织化学分析也证实了这些趋势。 Northern印迹分析表明，在2周龄的所有基因型中均检测到相当数量的OPN转录物，但仅在16周龄的twy小鼠中保持了OPN mRNA的强表达。无论16周大时的基因型如何，都几乎未检测到COL10A1转录本。结论：OPN在twy小鼠肥大性脊柱病变中过度表达，而肥大性增生主要是由于脊髓韧带异位骨化所致。由于OPN被认为是钙化的抑制剂，因此有必要进行进一步的研究以验证twy小鼠中OPN是否过表达。

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