DNA LINKAGE STUDY OF CLEFT LIP AND PALATE

这个项目的总体目标是描述分子机制。 遗传性非综合征性唇裂伴或不伴腭裂(CLP)。 面裂的原因是非常不同的，CLP是 超过100个可识别的症状的一部分，但更常见的是， 以孤立畸形的形式出现的。在后一组中 我们最近发现了一种称为非综合征性CLP的疾病 异质性，但已经确定了一组同质的 常染色体显性遗传性CLP多代家系。这些 多代家庭的特点很好，这是关键的一步 为阐明其分子机制及鉴定奠定基础 基因(S)负责。在最初的研究中，我们已经表明 常染色体显性遗传基因与部分CLP和 排除了中电与转化生长因子α的紧密联系 此前有报道称与可遗传的中电有关。这 建议为继续我们的研究提供资金， 常染色体显性遗传性唇腭裂 来自NIDR的小额赠款(R03)。在这个持续的项目中，我们建议 使用两种方法定位导致非综合征性CLP的一个(或多个)常染色体位点 “候选基因”和“反向遗传学”方法。我们正处于一个独特的 实现这一目标的位置，因为我们已经确定了同质 常染色体显性遗传性CLP多代家系，分子 定位基因座(Loci)的技术是可用的，并且有许多 生物学上相关的候选基因是可用的。 动物研究最近为颅面提供了新的见解 胚胎发生和细胞-细胞相互作用的作用。从这些 观察到，14个候选基因已经被挑选出来进行研究 南方凝胶方法学。受影响的限制片段模式 将对个体进行全面的染色体重排检查，如 作为删除或插入。将测试到多态站点的链接 在多代家族中确定候选基因是否 都与中电紧密相连。如果排除紧密联系，则形成 将用于构建连锁排除图谱。最后，如果 排除了与候选基因的连锁，对 基因组将使用数量可变的串联重复探针进行， 多酶切位点多态与聚合酶链式反应 可检测到的多态以检测与CLP基因座的连锁。被排除在外 从以前的连锁研究中开发的图谱和本研究中排除的基因座 研究将用于有针对性的研究领域。信息源自 这项研究将为1)阐明具体的 在这些多代家庭中导致遗传性慢性前列腺炎的突变(S)， 2)描述疾病的发病机制，以及，3)提供 为中电的高危家庭提供具体的咨询信息。 有关可遗传的中电的信息也可能有助于理解 散发性非综合征性CLP及其他疾病的发病机制 胚胎发育异常引起的疾病。

The overall aim of this project is to delineate the molecular mechanisms of heritable nonsyndromic cleft lip with or without cleft palate (CLP). The causes of facial clefting are extremely heterogeneous with CLP being part of greater than 100 recognizable syndromes but, more commonly, occurring as an isolated malformation. Within the latter group of disorders, called nonsyndromic CLP, we have recently shown genetic heterogeneity but have identified a homogeneous group of multigenerational families with autosomal dominant CLP. These multigenerational families have been well characterized, a crucial step towards elucidating the molecular mechanisms and the identification of the gene(s) responsible. In the initial study, we have shown that autosomal dominant gene is responsible for some of the cases of CLP and excluded tight linkage of CLP to transforming growth factor alpha which had previously been reported to be associated with heritable CLP. This proposal is for funds to continue our studies characterizing an autosomal dominant cleft lip and palate locus previously supported by a small grant (RO3) from the NIDR. In this continuing project, we propose to map an autosomal locus (or loci) causing nonsyndromic CLP using both "candidate gene" and "reverse genetics" approaches. We are in a unique position to accomplish this goal because we have identified homogeneous multigenerational families with autosomal dominant CLP, the molecular technology to localize the locus (loci) is available and a number of biologically pertinent candidate genes are available. Animal studies have recently provided new insights into craniofacial embryogenesis and the role of cell-cell interactions. From these observations, fourteen candidate genes have been selected for study by Sourthern gel methodology. Restriction fragment patterns from affected individuals will be examined for gross chromosomal rearrangements, such as deletions or insertions. Linkage to polymorphic sites will be tested in the multigenerational families to determine whether candidate genes are tightly linked to CLP. If tight linkage is excluded, the formation will be used in constructing linkage exclusion maps. Finally, if linkage to the candidate genes is excluded, a systemic search of the genome will be undertaken using variable number tandem repeat probes, multiple restriction site polymorphisms, and polymerase chain reaction detectable polymorphisms to detect linkage to a CLP locus. An exclusion map developed from previous linkage studies and loci excluded in this study will be used to target areas for study. Information derived from this study will provide a basis for 1) elucidating the specific mutation(s) causing heritable CLP in these multigenerational families, 2) characterizing the pathogenesis of the disorder and, 3) providing specific counseling information for families at risk for CLP. Information about heritable CLP may also lead to an understanding of the etiologic mechanisms involved in sporadic nonsyndromic CLP and other disorders resulting from abnormalities of embryogenesis.