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Function of GDFs in Tooth Development

GDF 在牙齿发育中的功能

基本信息

批准号：
09044320
负责人：
AKAMINE Akifumi
金额：
$ 3.2万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044320/
关键词：
Gdf11 BMP tooth morphogenesis dental pulp gene targeting Gli zinc finger transcription factor defferentiation factor Growth differetiation factor ノックアウトマウス zinc finger transcription factor 歯髄歯牙発生 Growth differentiation factor

项目摘要

We have cloned and characterized a new member of the BMP/TGFb superfamily, Gdf 11, from rat dental(incisor) pulp RNA by RT-PCR using degenerate primers. The mature carboxyl-terminal domain encoded by Gdf11 is 85% identical to mouse Growth/differentiation factor 8 (Gdf8). Northern blot analysis revealed Gdf11 is expressed in adult dental pulp and brain. In situ hybridization of sections and whole mount embryos demonstrated Gdf 11 is expressed as early as 8.5 days post coitus (dpc) in the tail bud. At 10.5 dpc, it is expressed in the branchial arches, limb bud, tail bud and posterior dorsal neural tube. Later, it is expressed in terminaliy differentiated odontoblasts, the nasal epithelium, retina and specific regions of the brain. These studies indicate that Gdf11 could cooperate with other BMP/TGFb superfamily members in multiple developmental processes. The mice lack of Gdf 11, however, did not have phenotype.We have also cloned a new member of the zinc finger transcription factor, G23 … More from incisor pulp RNA by RT-PCR using degenerate primers. The five zinc finger domain encoded by mouse G23 has almost 65 % amino acid sequence homology with Gli1, Gli2 and Gli3. Northern blot analysis revealed G23 is expressed in dental pulp, kidney and testis. In situ hybridization of sections and whole mount embryos demonstrated G23 is first detected diffusely in forelimb and hindlimb bud at 10.0 days post coitus (dpc). At 10.5 dpc, it is expressed in the branchial arches, limb bud, craniofacial border, and ventral part of the tail. Later, it is expressed in whisker follicle, intervertebral disc, kidney, and testis. The expression of G23 was compared with the genes involved in the Sonic Hedgehog (Shh) signaling pathway, Shh, Ptc, Gui1, Gli2, and Gli3 during tooth development. G23 was found to be expressed in dental papillae in temporally and spatially overlapped pattern with Ptc, Glil, Gli2, and Gli3 where Shh are not expressed. In order to assess the possible role of G23 in Shh signaling pathway and its interaction with Glis, the expression of G23 in Glil/Gli2 mutant and Shh mutant was examined during tooth development and kidney development. The G23 transcript was upregulated in tooth germ in Gli2-/- and Glil+/- ; Gli2-/- mutant, suggesting G23 might have a role of suppresser on the Shh signaling pathway during tooth development. The expression of G23 in kidney was not changed in Gli2-/- and Glil+/- ; Gli2-/- mutant. There might be different feedback pathways operating to regulate the action of Shh in the cell proliferation and maintenance between tooth and kidney development. G23 might have redundant function and display similar signaling activity with Gli members. To examine more directly the role of G23 in embryogenesis, special reference to tooth and kidney development, the targeted gene disruption of G23 in the mouse was performed. Less

我们使用简并引物通过 RT-PCR 从大鼠牙（切牙）牙髓 RNA 中克隆并表征了 BMP/TGFb 超家族的新成员 Gdf 11。 Gdf11 编码的成熟羧基末端结构域与小鼠生长/分化因子 8 (Gdf8) 85% 相同。 Northern blot 分析显示 Gdf11 在成人牙髓和大脑中表达。切片和整体胚胎的原位杂交证明 Gdf 11 早在性交后 8.5 天 (dpc) 就在尾芽中表达。在 10.5 dpc 时，它在鳃弓、肢芽、尾芽和后背神经管中表达。随后，它在终末分化的成牙本质细胞、鼻上皮、视网膜和大脑的特定区域中表达。这些研究表明，Gdf11 可以在多个发育过程中与其他 BMP/TGFb 超家族成员合作。然而，缺乏Gdf 11的小鼠没有表型。我们还使用简并引物通过RT-PCR从门牙髓RNA中克隆了锌指转录因子的新成员G23…更多。小鼠G23编码的五个锌指结构域与Gli1、Gli2和Gli3具有近65%的氨基酸序列同源性。 Northern印迹分析显示G23在牙髓、肾脏和睾丸中表达。切片和整体胚胎的原位杂交表明，在性交后 10 天 (dpc) 时，首先在前肢和后肢芽中广泛检测到 G23。在 10.5 dpc 时，它在鳃弓、肢芽、颅面边缘和尾部腹侧部分表达。随后，它在须毛囊、椎间盘、肾脏和睾丸中表达。将 G23 的表达与牙齿发育过程中涉及 Sonic Hedgehog (Shh) 信号通路的基因 Shh、Ptc、Gui1、Gli2 和 Gli3 进行比较。发现 G23 在牙乳头中以与 Ptc、Glil、Gli2 和 Gli3 在时间和空间上重叠的模式表达，而 Shh 不表达。为了评估G23在Shh信号通路中的可能作用及其与Glis的相互作用，检测了Glil/Gli2突变体和Shh突变体在牙齿发育和肾脏发育过程中G23的表达。 Gli2-/- 和 Glil+/- 牙胚中 G23 转录本上调； Gli2-/- 突变体，表明 G23 可能在牙齿发育过程中对 Shh 信号通路具有抑制作用。肾脏中G23的表达在Gli2-/-和Glil+/-中没有变化； Gli2-/- 突变体。可能存在不同的反馈途径来调节 Shh 在牙齿和肾脏发育之间的细胞增殖和维持中的作用。 G23 可能具有冗余功能并显示与 Gli 成员相似的信号活动。为了更直接地检查 G23 在胚胎发生中的作用，特别是牙齿和肾脏发育，在小鼠中进行了 G23 的靶向基因破坏。较少的