A Rapid, Nonradioactive In Situ Hybridization Technique for Use on Cryosectioned Adult Mouse Bone
A Rapid, Nonradioactive In Situ Hybridization Technique for Use on Cryosectioned Adult Mouse Bone
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DOI：
10.1007/s00223-008-9154-1
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发表时间：
2008-09-01
影响因子：
4.2
通讯作者：
Susa, Mira
中科院分区：
文献类型：
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作者：
Salie, Rishard;Li, Haitao;Susa, Mira
成人骨的原位杂交（ISH）是一项困难的任务，需要至少3-5周的脱钙、石蜡包埋和切片。由于这个原因，骨ISH通常只在胚胎或新生动物组织上进行，而成人中的基因表达问题尚未得到解决。在这里，我们报告的发展ISH系统，只需要7天的无酸脱钙，包埋，切片，有利于保存组织mRNA的条件。使用CryoJane Tape Transfer系统切割来自3-12周龄成年小鼠的组织冷冻切片。石蜡切片和冷冻切片的组织具有相当的形态学，并且用地高辛标记的核糖核酸探针对在骨形成成骨细胞（I型胶原，骨钙素，Runx 2）或肥大或增殖软骨细胞（X型胶原，Runx 2）中发现的mRNA进行酶染色。
In situ hybridization (ISH) of adult bone is a difficult task that requires at least 3-5 weeks for decalcification, paraffin embedding, and sectioning. For that reason, bone ISH is often done only on embryonic or newborn animal tissue, leaving unanswered the question of gene expression in adults. Here, we report the development of an ISH system that requires only 7 days for acid-free decalcifiation, embedding, and sectioning, conditions that are conducive to preservation of tissue mRNA. The tissue cryosections, derived from adult mice 3-12 weeks old, were cut using the CryoJane Tape Transfer system. Paraffin-sectioned and cryosectioned tissue have comparable morphology and stained enzymatically with digoxigenin-labeled riboprobes for mRNA found in either bone-forming osteoblasts (type I collagen, osteocalcin, Runx2) or the hypertrophic or proliferating chondrocytes (type X collagen, Runx2).