用原位杂交技术测定MMP13mRNA在小鼠长骨发育中的表达

Detection of MMP13 mRNA expression in mouse long bone during the development using in situ hybridization technique

目的通过测定MMP13mRNA在小鼠长骨发育不同阶段的表达,进一步证实MMP13的表达位置及作用,通过其mRNA表达的信号强弱变化,进一步探讨MMP13在长骨发育中的作用。方法应用原位杂交技术,在小鼠不同发育阶段的长骨组织切片中,用同位素标记探针显示MMP13mRNA的基因表达。结果在小鼠胚胎15.5天、17.5天、出生第一天、两周及一月,均可见在生长带软骨及软骨与骨交界处,有大量MMP13的基因表达,并随着长骨发育的成熟,其表达的信号强度逐渐下降。结论 MMP13在胚胎发育过程中,在长骨生长带的肥大软骨细胞及与原发性骨化中心中有基因表达,在胚胎发育中,由于发育快,基因表达非常强烈,随着长骨发育的成熟,MMP13mRNA表达逐渐减弱,证实其在长骨的发育中起到重要的作用。同时,此研究表明原位杂交技术用同位素标记显像的方法,在组织切片中测定基因的表达直观准确,值得推广应用。

Objective To determine the expression and location of MMP13 mRNA in the long bone during the different period of the development in mice, and to explore the effect of MMP13 in the long bone development according to its mRNA signal expression level. Methods Using in situ hybridization technique, MMP13 mRNA expression was labeled with isotopic probe on histological sections from the long bone in different period of development in mice. Results High level of MMP13 expression were seen in the end of the growth plate and primary spongiosa on El5.5, El7.5, 1 day, 2 weeks, and 1 month after birth of the mice. With the mature of the long bone, the expression level decreased gradually. Conclusion MMP13 is expressed in hypertrophic chondrocytes in the end of the growth plate and osteoblasts in the primary spongiosa of the long bone during the embryonic development. Because of the fast development, MMP13 expression is very strong during the period of embryo. Following the mature of the long bone, the expression was decreased gradually, indicating that MMP13 plays an important role in the development of long bone. Meanwhile, this study demonstrates that in situ hybridization technique using isotopic labeling and imaging is a straight and accurate method for measurement of gene expression in tissue sections, and should be applied broadly.