DEVELOPMENT OF PALATE AND CRANIOFACIAL MESENCHYME

腭和颅面间充质的发育

• 批准号: 2132276
• 负责人: ELIZABETH D HAY
• 金额: $ 21.95万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2132276
• 关键词: Retroviridae; antibody; antisense nucleic acid; cell differentiation; cell transformation; chick embryo; cleft palate; connective tissue development; embryo /fetus tissue /cell culture; embryogenesis; epithelium; face; head; histogenesis; immunocytochemistry; in situ hybridization; laboratory mouse; laboratory rat; mesenchyme; neural crest; oligonucleotides; osteocytes; palate; transforming growth factors; transmission electron microscopy

This grant proposal is about the development of the palate, with special attention to the role of the medial edge epithelium (MEE) and cranial neural crest mesenchyme. Mesenchymal cells arise by epithelial-mesenchymal transformation (BMT). They have the ability to invade collagen gels and other extracellular matrix (ECM), whereas epithelia sit on top of ECM and exhibit apical-basal polarity. Mesenchymal cells migrate through the matrix with front end/back end polarity. They differentiate into the cells that inhabit the ECM interior: fibroblasts, bone, melanocytes, and neurones. We will consider the following aspects of the origin and differentiation of the craniofacial mesenchymes that contribute to palatal morphogenesis. I. Mesenchymal Cell Lineages in the Developing Palate. Using recombinant retroviruses to label specific groups of cells in the embryo, we expect to confirm and extend our earlier study that MEE-derived mesenchyme forms fibroblasts but not bone, and to prove for the first time that head neural crest in the rodent consists of two physically separate cell populations, one of which forms connective tissue and the other, melanocytes and neurones. The significance of this study to students of palate morphogenesis is that it establishes a multiple origin of palate mesenchyme and calls attention to the fact that palate MEE does not die as previous authors thought. It casts a new light on roles the MEE might play in cleft palate. II. Effect of Bone Inducing Molecules on the Differentiation of Mesenchymal Cell Lineages. We will attempt to drive the cells above that do not normally form cartilage/bone to do so by administering BMP to them in vitro. It will be particularly interesting to see if mesenchyme formed from palate epithelium and neurogenic crest cells will undergo chondrogenesis under the influence of this powerful inducing agent. The results could have important impact on our ideas about the ability of "determined" cells (e.g., neurogenic crest) to reawaken apparently closed pathways of differentiation. III. Induction and Inhibition of Epithelial Fusion and EMT in Nonfusing and Fusing Palates. Chicken palates do not normally fuse probably because the MEE stratifies and periderm does not slough to expose the basal epithelia to each other. We will surgically remove the periderm and place chicken palatal shelves in close contact to try to induce formation of a midline seam. TGFbeta3 will be added to promote EMT from this seam and the fate of mesenchymal cells followed. Antibodies and antisense oligonucleotides to TGFbeta1-3 will be added to try to inhibit EMT from MEE of fused rodent palates. The recognition our recent work has given to the fact that MEE transforms to mesenchyme establishes a whole new way of looking at the possible role of TGFbeta's and sloughing of the outer layer of the MEE in palatogenesis. The immediate health relevance is to understanding normal development, but the work could give new insights into causes of congenital anomalies, especially cleft palate.

该赠款提案是关于味蕾的发展，特别是 注意内侧边缘上皮（MEE）和颅骨的作用 神经rest间充质。间充质细胞由上皮 - 间质产生 转换（BMT）。他们有能力侵入胶原蛋白凝胶和 其他细胞外基质（ECM），而上皮位于ECM的顶部， 表现出顶端极性。间充质细胞通过 具有前端/后端极性的矩阵。它们分化为细胞 居住在ECM内部：成纤维细胞，骨骼，黑色素细胞和 神经元。我们将考虑来源的以下方面 颅面间质的分化，导致palatal 形态发生。 I.发育中的口感中的间充质细胞谱系。使用重组 逆转录病毒以标记胚胎中特定细胞的特定组，我们期望 确认并扩展了我们较早的研究表明MEE衍生的间质形成 成纤维细胞但不是骨头，并首次证明头神经 啮齿动物中的波峰由两个物理分开的细胞种群组成， 其中一种形成结缔组织，另一种是黑素细胞和 神经元。这项研究对味觉学生的意义 形态发生是它建立了pa的多重来源 间充质并引起人们的注意，即papate mee不会死于 以前的作者认为。它对MEE可能扮演的角色投入了新的启示 在left裂中。 ii。骨骼诱导分子对分化的影响 间充质细胞谱系。我们将尝试驱动上方的细胞 通常不要通过向其施用BMP来形成软骨/骨骼 体外。看看是否形成间充质会特别有趣 从pa酸上皮和神经源性冠状细胞将经历 在这种强大的诱导剂的影响下，软骨发生。这 结果可能会对我们关于能力的想法产生重要影响 “确定”的细胞（例如，神经源性波峰）显然关闭了 分化途径。 iii。在非诱导中诱导和抑制上皮融合和EMT 和融合味觉。鸡肉味通常不会融合，可能是因为 MEE分层，Periderm不会脱落以暴露基础 彼此上皮。我们将通过手术去除Periderm并放置 紧密接触的鸡palatal货架试图引起形成 中线接缝。将添加TGFBETA3，以促进此接缝的EMT和 伴随间充质细胞的命运。抗体和反义 将添加寡核苷酸到TGFBETA1-3，以尝试抑制EMT 融合啮齿动物的味觉。我们最近的工作给予了认可 MEE转变为间充质的事实建立了一种全新的方式 查看TGFBETA的可能作用和外层的链条 在古质发生中的mee。 直接健康的相关性是理解正常发展，但 这项工作可以给予新的见解，以了解先天性异常的原因， 尤其是口味。