Regulation and Functions of Adamts9 During Ovulation

Adamts9在排卵期间的调节和功能

• 批准号: 9303524
• 负责人: Yong Zhu
• 金额: $ 42.62万
• 依托单位: EAST CAROLINA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9303524
• 关键词: ADAMTS1 gene; ADAMTS9 gene; Affect; Anovulation; CRISPR/Cas technology; Cells; Data; Development; Disease; Disintegrins; Drug Targeting; Enzyme Activation; Enzymes; Event; Extracellular Matrix; Family; Fertility; Fishes; Foundations; Future; Gene Expression; Gene Targeting; Genes; Gonadotropins; Heat-Shock Response; Hormonal; Human; Infertility; Knock-out; Knockout Mice; Malignant Neoplasms; Mammals; Matrix Metalloproteinases; Mediating; Mental disorders; Metalloproteases; Modeling; Molecular; Nuclear; Oocytes; Outcome; Ovulation; Peptide Hydrolases; Pharmacotherapy; Phenotype; Process; Progesterone Receptors; Progestins; Proteins; Rattus; Regulation; Resources; Rupture; Signal Pathway; Signal Transduction; Signaling Protein; Steroid Receptors; Steroids; Thrombospondins; Time; Tissue-Specific Gene Expression; Tissues; Transcript; Zebrafish; experimental study; genome-wide; hormone regulation; infertility treatment; member; new therapeutic target; novel; overexpression; ovulation disorders; reproductive; transcription factor; transcriptome sequencing

Ovulation is an indispensable reproductive event; yet our understanding of the molecular mechanisms that control ovulation is still incomplete. Activation of proteases leading to extracellular matrix disassociation is an essential step for follicle rupture and ovulation to proceed. Involvement of several metalloproteinase families including members of ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs), MMPs (matrix metalloproteinases), and ADAM (a disintegrin and metalloproteinase) in ovulation and tissue remodeling has been suggested. However, the molecular mechanisms regulating key proteinases and identities of essential enzymes that are responsible for the recurring tissue remodeling process have not been established. Our preliminary data suggest that a member of ADAMTS family, ADAMTS9, is likely a downstream target of nuclear progestin receptor (PGR), and is mainly responsible for ovulation. We will use CRISPR/Cas9 to generate global and conditional knockouts of ADAMTS9, and characterize these knockouts to determine whether knockout ADAMTS9 affects ovulation and fertility in zebrafish. If ADAMTS9 is a key downstream enzyme of PGR that is important for ovulation, we should be able to rescue PGR knockout anovulation phenotype by overexpressing ADAMTS9 in PGR knockout fish. In addition, we propose to determine the expression, regulation and functions of ADAMTS9 during ovulation in zebrafish, in order to elucidate the molecular mechanisms and signaling pathways for ovulation. We will determine expression of both transcript and protein of ADAMTS9 during the development of oocytes, and in the follicular cells of preovulatory oocytes around the ovulation in zebrafish. We will also determine whether the expression of ADAMTS9 is hormonally regulated by gonadotropins and/or progestin. We will then determine whether PGR regulates ADAMTS9 directly or indirectly via other transcriptional factors. For the first time, the regulation and functions of ADAMTS9, particularly in the follicular cells of oocytes, will be comprehensively determined in a vertebrate model. The ADAMTS9 knockout models will be very valuable resource for future studies. The functions of ADAMTS9 in ovulation and fertility will also be established in a zebrafish model, which will provide a foundation to study the functions and regulation of ADAMTS9 in mammals, as well as the development of new drug targets and novel non-steroid treatments for infertility, tissue remodeling, and cancer.

排卵是必不可少的生殖事件；然而，我们对分子的理解 控制排卵的机制仍然不完整。激活蛋白酶导致 细胞外基质分离是卵泡破裂和排卵进行的重要步骤。 包括Adamts成员在内的几个金属蛋白酶家族的参与（崩解蛋白 具有血小板蛋白基序），MMP（基质金属蛋白酶）和Adam的金属蛋白酶酶 （一种崩解蛋白和金属蛋白酶）在排卵和组织重塑中已提出。 但是，调节关键蛋白酶的分子机制和必需的身份 负责重复组织重塑过程的酶尚未 已确立的。我们的初步数据表明，Adamts家族的成员Adamts9可能是 核孕激素受体（PGR）的下游靶标，主要负责排卵。 我们将使用CRISPR/CAS9生成ADAMTS9的全球和有条件淘汰 表征这些敲除以确定敲除adamts9是否影响排卵和 斑马鱼的生育能力。如果ADAMTS9是PGR下游酶的关键，这对于 排卵，我们应该能够通过过表达来挽救PGR敲除表型 Adamts9在PGR淘汰鱼类中。此外，我们建议确定表达式，调节 斑马鱼排卵期间Adamts9的功能，以阐明分子 排卵的机理和信号通路。我们将确定这两个笔录的表达 和ADAMTS9的蛋白质在卵母细胞发展过程中以及在卵泡细胞中 斑马鱼排卵周围的排卵前卵母细胞。我们还将确定是否 ADAMTS9的表达受促性腺激素和/或孕激素的荷尔蒙调节。然后我们会 确定PGR是否通过其他转录因子直接或间接调节ADAMTS9。 第一次，ADAMTS9的调节和功能，特别是在 卵母细胞将在脊椎动物模型中全面确定。 Adamts9淘汰赛 对于将来的研究，模型将是非常宝贵的资源。 ADAMTS9在排卵中的功能 斑马鱼模型还将建立生育能力，该模型将为研究提供基础 哺乳动物中ADAMTS9的功能和调节以及新药的发展 靶标和新型的非甾体类药物治疗，用于不育，组织重塑和癌症。