DEFECTS IN MULLERIAN DUCT REGRESSION

苗勒管回归缺陷

• 批准号: 3316646
• 负责人: VICKI N MEYERS-WALLEN
• 金额: $ 15.27万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1992-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3316646
• 关键词: Mullerian duct inhibiting substance; cytogenetics; dogs; electron microscopy; embryo /fetus cell /tissue; endocrine disorder; fallopian tubes; genetic regulation; genital secretion; gonads; hermaphroditism; histology; messenger RNA; microscopy; morphology; northern blottings; organ culture; pregnancy; pseudohermaphroditism; sex chromosomes; sex development disorder; sex differentiation; sex linked trait; testis; testis disorder

In two separate hereditary syndromes in the dog, Mullerian ducts persist in the presence of testicular tissue: XX sex reversal and the persistent Mullerian Duct Syndrome (PMDS). These are the only known animal models with consistent and specific inherited defects in repression of the Mullerian duct system described to date. Preservation of these models for further study depends entirely upon this proposal, since there is no other funding to maintain these animals. Their significance is not only that they are models of human developmental disorders. They are an important resource for understanding the role of Mullerian Inhibiting Substance (MIS) in mammalian reproductive development and function. Although the gene for MIS has been cloned and biochemical mechanisms of its action have been proposed, the MIS receptor has not been identified. These models should lead to a more detailed understanding of the genetic control, structure, and mechanisms of action of MIS. In both syndromes, we have shown that failure of Mullerian regression is unlikely to result from an absence of MIS since we have recently shown that testicular tissue of affected dogs causes regression of embryonic rat Mullerian ducts in an organ culture bioassay. Our objective in the present study is to determine whether the apparent insensitivity of the Mullerian system to MIS is due to a defect in timing or amount of MIS. secretion or to a defect in the MIS receptor. These hypotheses will be tested by: 1) Determine whether the synthesis and timing of MIS mRNA and MIS in affected dogs is comparable to that in normal male littermates, and 2) Comparing the location, concentration, and MIS binding characteristics of the MIS receptor in affected dogs and normal littermates. Dogs with XX sex reversal (Cocker Spaniels) and PMDS (Miniature Schnauzers) will be produced by breeding known carriers of these autosomal recessive traits. Production and cellular location of MIS mRNA in testes of normal and affected littermates will be quantitated and compared by Northern blot, in situ hybridization, and immunohistochemical techniques. MIS levels in testes of normal and affected littermates will be determined by Western blot analysis. The MlS receptor will first be identified by antiidiotypic antibodies and subsequently by binding to metabolically-labelled MIS produced by stable cell lines transfected with the recombinant human MIS gene.

在两个独立的遗传综合征的狗，苗勒管持续存在， 睾丸组织的存在：XX性反转和持续性 苗勒管综合征（PMDS）。 这些是唯一已知的动物模型 具有一致的和特异性的遗传缺陷， 迄今为止描述的苗勒管系统。 保存这些模型， 进一步的研究完全取决于这一建议，因为没有其他的建议。 为这些动物提供资金。 它们的意义不仅在于此 它们是人类发育障碍的模型。 他们是一个重要的 了解苗勒管抑制物质（MIS）的作用的资源 哺乳动物生殖发育和功能。 虽然基因为 MIS已被克隆，其作用的生化机制已被证实。 提出，MIS受体尚未确定。 这些模型应该 导致对遗传控制，结构， 和MIS的作用机制。 在这两种综合症中，我们已经表明， 缪勒回归的失败不太可能是由于缺乏 因为我们最近发现，受影响的狗的睾丸组织 导致胚胎大鼠苗勒管在器官培养中退化 生物测定 本研究的目的是确定 苗勒系统对MIS的明显不敏感是由于 的时间或数量。 分泌或MIS受体的缺陷。 这些假设将通过以下方式进行检验：1）确定综合和 受影响犬MIS mRNA和MIS的时间与正常犬相当 雄性同窝仔，以及2）比较位置、浓度和MIS MIS受体在患病犬和正常犬中的结合特征 同窝出生的 XX性反转犬（可卡犬）和PMDS（小型雪纳瑞） 将通过繁殖已知的这些常染色体隐性遗传的携带者产生 性状 正常人睾丸MIS mRNA的产生及其细胞定位 并通过北方印迹对受影响的同窝仔进行定量和比较， 原位杂交和免疫组织化学技术。 管理信息系统水平 正常和受影响同窝仔的睾丸将由Western 印迹分析 MlS受体首先将通过抗独特型抗体 随后通过结合代谢标记的MIS 由重组人MIS转染的稳定细胞系产生 基因