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Analysis of the function of PTHrP in tooth development.

PTHrP在牙齿发育中的功能分析

基本信息

批准号：
08457479
负责人：
KURISU Kojiro
金额：
$ 5.5万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457479/
关键词：
Tooth germ PTHrP Organ culture Mouse Morphogenesis Development Tissue differentiation Antisense oligonucleotide

项目摘要

Gene expression of PTHrP was observed in the enamel organ of rat tooth germ, whereas that of its receptor, PTH/PTHrP receptor, was in the dental papilla and surrounding mesenchyme. When the mouse tooth explants were cultured with antisense oligodeoxynucleotide (ODN) against mouse PTHrP mRNA in serum-free medium, the invasion of bone tissue was observed in the tooth germs as follows. At 5-6 days in culture, the bone tissue extending from outside of tooth germ began to invade into the dental papilla. The invasion of bone tissue into developed extensively during the culture period and the invasion occurred also into enamel organ. During 10-21 days in culture the tooth germs were destroyed by the extensive invasion of bone tissue. On the other hand, the explants cultured without ODN,with sense ODN or with antisense ODN plus recombinant human PTHrP showed normal histological structure as observed in vivo. Antisense ODN for mouse PTH/PTHrP receptor also induced the bone invasion into the tooth germs as observed in the explants treated with antisense ODN for PTHrP.The treatment with antisense ODN for PTHrP induced the reduction of TRAP (tartrate-resistant acid phophatase) activity in osteoclasts in the explants and suppressed the formation of dental sac, sugesting that the reductions in osteoclast activity and in dental sac formation are involved in the bone invasion induced the antisense-treatment.Results suggest that PTHrP is essential for the tooth development and for the protection of tooth germs from the invasion of bone tissues.

PTHrP基因在大鼠牙胚的牙釉质器官中表达，而其受体PTH/PTHrP受体在牙乳头和周围间质中表达。用抗小鼠PTHrP mRNA的反义寡脱氧核苷酸（ODN）在无血清培养基中培养小鼠牙胚，观察牙胚对骨组织的侵袭情况。培养5-6天后，牙胚外延伸的骨组织开始侵入牙乳头。在培养过程中，骨组织对牙釉质器官的侵袭十分普遍。在10-21天的培养过程中，牙胚被广泛侵入的骨组织破坏。另一方面，无ODN、带正义ODN和带反义ODN +重组人PTHrP的外植体在体内的组织结构均正常。小鼠PTH/PTHrP受体的反义ODN在PTHrP的反义ODN处理的外植体中也能诱导骨侵入牙胚。反义ODN处理PTHrP诱导外植体破骨细胞TRAP（酒石酸抗性酸性磷酸酶）活性降低，抑制牙囊形成，提示破骨细胞活性降低和牙囊形成减少参与了反义处理诱导的骨侵。结果表明，PTHrP对牙齿发育和保护牙齿细菌免受骨组织的侵害至关重要。