Role of Indian hedgehog signaling in palatal osteogenesis.
Role of Indian hedgehog signaling in palatal osteogenesis.
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DOI：
10.1097/prs.0b013e3182043a07
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发表时间：
2011-03
影响因子：
3.6
通讯作者：
Longaker MT
中科院分区：
文献类型：
--
作者：
Levi B;James AW;Nelson ER;Brugmann SA;Sorkin M;Manu A;Longaker MT
唇裂和腭裂是常见的出生缺陷，代表着巨大的生物医学负担。通过使用动物模型，腭裂的分子基础变得越来越清晰。 Indian Hedgheg (Ihh) 已被证明与颅面发育相关并且在腭骨中活跃。我们假设 Ihh 活性在次级腭内的成骨中发挥作用，并且该途径的缺陷可能会抑制次级腭的成骨。在上颚发育期间(e9.5-e17.5)从野生型小鼠中分离出上颚。 qRT-PCR用于检测成骨分化和细胞分化过程中的基因表达（Shh、Ihh、Ptc1、Gli1、Gli2、Gli3、Runx2、Alp、Col1a1）。接下来，通过 H&E、苯胺蓝、Pentachrome 和原位杂交对腭进行分析，以评估腭架的成骨和刺猬通路基因的表达。最后，对印度刺猬缺失小鼠的上颚进行了分析，以确定遗传缺陷对上颚发育成骨的影响。印度刺猬和成骨信号的增加与野生型小鼠上颚的骨化和融合同时发生。与 e9.5 相比，这包括在 e15.5 时 Hedgehog 元件（包括 Ihh）的表达峰值达到 5-150 倍。相反，由于基因敲除（Ihh -/-）而导致印度刺猬的损失导致继发性腭骨化减少。我们的结果表明 Hedgehog 信号在腭骨化过程中发挥作用。在腭融合过程中，刺猬通路被激活，而印度刺猬的缺失会导致次级硬腭的骨化减少。
Cleft lip and palate are common birth defects and represent a large biomedical burden. Through the use of animal models, the molecular underpinnings of cleft palate are becoming increasingly clear. Indian Hedgheg (Ihh) has been shown to be associated craniofacial development and active in the palatine bone. We hypothesize that Ihh activity plays a role in osteogenesis within the secondary palate and that defects in this pathway may inhibit the osteogenesis of the secondary palate. Palates were isolated from wild type mice during the period of palate development (e9.5-e17.5). qRT-PCR was used for detecting gene expression during osteogenic differentiation and cellular differentiation (Shh, Ihh, Ptc1, Gli1, Gli2, Gli3, Runx2, Alp, Col1a1). Next, palates were analyzed by H&E, Aniline Blue, Pentachrome, and In situ Hybridization to assess osteogenesis of the palatal shelf and expression of hedgehog pathway genes. Finally, the palate of Indian Hedgehog null mice was analyzed to determine the effect of genetic deficiency on palatal development osteogenesis. Increased Indian Hedgehog and osteogenic signaling coincided with ossification and fusion of the palate in wild-type mice. This included a 5-150 fold peak in expression of Hedgehog elements, including Ihh, at e15.5 as compared to e9.5. Contrarily, loss of Indian Hedgehog by genetic knockout (Ihh −/−) resulted in decreased secondary palate ossification. Our results suggest a role for Hedgehog signaling during palatal ossification. The hedgehog pathway is activated during palatal fusion and deletion of Indian hedgehog leads to diminished ossification of the secondary hard palate.