牙本质涎磷蛋白、Ⅰ型胶原蛋白在vps4b基因敲除鼠磨牙牙胚发育中的时空表达

doi:10.7518/hxkq.2019.03.004

华西口腔医学杂志 ›› 2019, Vol. 37 ›› Issue (3): 248-252.doi: 10.7518/hxkq.2019.03.004

牙本质涎磷蛋白、Ⅰ型胶原蛋白在vps4b基因敲除鼠磨牙牙胚发育中的时空表达

陈栋¹(),王莹莹¹,李晓聪²,鲁方丽¹,李强¹

1.郑州大学第一附属医院口腔科,郑州 450000
2.濮阳市人民医院口腔科,濮阳 457099

收稿日期:2018-12-23 修回日期:2019-03-19 出版日期:2019-06-01 发布日期:2019-06-12
通讯作者: 陈栋 E-mail:chendongfmmu@163.com
作者简介:陈栋,教授,博士,E-mail:chendongfmmu@163.com
基金资助:
国家自然科学基金(81470033)

Spatio-temporal expression of dentin sialophosphoprotein and collagen Ⅰ during molar tooth germ development in vps4b knockout mouse

Dong Chen¹(),Yingying Wang¹,Xiaocong Li²,Fangli Lu¹,Qiang Li¹

1.Dept. of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, China
2.Dept. of Stomatology, Puyang People’s Hospital, Puyang 457099, China

Received:2018-12-23 Revised:2019-03-19 Online:2019-06-01 Published:2019-06-12
Contact: Dong Chen E-mail:chendongfmmu@163.com
Supported by:
The National Natural Science Foundation of China(81470033)

摘要/Abstract

摘要：

目的研究vps4b基因突变对牙齿发育相关蛋白——牙本质涎磷蛋白（DSPP）和Ⅰ型胶原蛋白（COL-Ⅰ）表达的影响。方法取胚胎E13.5 d、E14.5 d、E16.5 d的胎鼠头部及出生后P2.5 d、P7 d的幼鼠下颌骨组织,石蜡包埋后获取第一磨牙牙胚组织切片,采用免疫组织化学染色法检测野生型小鼠和vps4b基因敲除小鼠牙胚中DSPP、COL-Ⅰ的表达。结果野生鼠蕾状期和帽状期DSPP、COL-Ⅰ均未表达;钟状期DSPP在内釉上皮和牙乳头中有表达,COL-Ⅰ表达于牙乳头和牙囊;分泌期和矿化期DSPP、COL-Ⅰ在成釉细胞、成牙本质细胞、牙囊内均有表达,COL-Ⅰ在牙乳头内亦可见表达。小鼠vps4b基因敲除后,DSPP在钟状期牙乳头和分泌期牙乳头及牙囊内未见表达,COL-Ⅰ在钟状期及矿化期表达部位与野生型小鼠一致,分泌期在牙乳头的表达发生改变。结论 vps4b基因在牙胚发育中发挥重要作用;DSPP、COL-Ⅰ的表达可能受vps4b基因的调控,并与vps4b共同调节牙齿牙本质的发育。

关键词: 牙本质涎磷蛋白, Ⅰ型胶原蛋白;, vps4b基因敲除鼠, 牙胚发育

Abstract:

Objective To verify the effect of the mutant gene vps4b on the expression of tooth development-related proteins, dentin sialophosphoprotein (DSPP) and collagenⅠ (COL-Ⅰ). Methods Paraffin tissue sections of the first molar tooth germ were obtained from the heads of fetal mice at the embryonic stages of 13.5, 14.5, and 16.5 days and from the mandibles of larvae aged 2.5 and 7 days after birth. The immunohistochemical method was used to detect the expression and location of DSPP and COL-Ⅰ in wild-type mouse and vps4b knockout mouse. Results DSPP and COL-Ⅰ were not found in the bud and cap stages of wild-type mouse molar germ. In the bell stage, DSPP was positively expressed in the inner enamel epithelium and dental papilla, whereas COL-Ⅰ was strongly expressed in the dental papilla and dental follicle. During the secretory and mineralized periods, DSPP and COL-Ⅰ were intensely observed in ameloblasts, odontoblasts, and dental follicles, but COL-Ⅰ was also expressed in the dental papilla. After vps4b gene knockout, DSPP was not expressed in the dental papilla of the bell stage and in the dental papilla and dental follicle of the secretory phase. The expression position of COL-Ⅰ in the bell and mineralization phase was consistent with that in the wild-type mice. Moreover, the expression of COL-Ⅰ in the dental papilla changed in the secretory stage. Conclusion Gene vps4b plays a significant role in the development of tooth germ. The expression of DSPP and COL-Ⅰ may be controlled by gene vps4b and regulates the development of tooth dentin and cementum together with vps4b.

Key words: dentin sialophosphoprotein, collagenⅠ, vps4b knockout mouse, tooth germ development

中图分类号:

R780.2

陈栋,王莹莹,李晓聪,鲁方丽,李强. 牙本质涎磷蛋白、Ⅰ型胶原蛋白在vps4b基因敲除鼠磨牙牙胚发育中的时空表达[J]. 华西口腔医学杂志, 2019, 37(3): 248-252.

Dong Chen,Yingying Wang,Xiaocong Li,Fangli Lu,Qiang Li. Spatio-temporal expression of dentin sialophosphoprotein and collagen Ⅰ during molar tooth germ development in vps4b knockout mouse[J]. West China Journal of Stomatology, 2019, 37(3): 248-252.

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基因	引物序列（5’—3’）
vps4b-1	F1:TGCTTTAGGCAAAGCAGCAC
	R1:GCTGAAACTGGAGGGTTAGG
vps4b-2	F2:TGCTTTAGGCAAAGCAGCAC
	R2:TTTCTGTGTGAGGGAGGCAA

牙本质涎磷蛋白、Ⅰ型胶原蛋白在vps4b基因敲除鼠磨牙牙胚发育中的时空表达

Spatio-temporal expression of dentin sialophosphoprotein and collagen Ⅰ during molar tooth germ development in vps4b knockout mouse

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