GLUTS AND GLUCOSE TRANSPORT IN THE MOUSE BLASTOCYST

小鼠胚泡中的过剩和葡萄糖运输

基本信息

批准号：
8119073
负责人：
KELLE H MOLEY
金额：
$ 24.56万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-02-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8119073
关键词：
Address Adult Affect Amino Acids Apoptosis Area Autophagocytosis Biochemical Cardiovascular Diseases Cell physiology Cells Competence Congenital Abnormality Data Defect Defense Mechanisms Development Developmental Delay Disorders Diabetes Mellitus Diabetic mouse Digestion Embryo Embryo Transfer Embryonic Development Equilibrium Etiology Event Exhibits Familiarity Fertilization Fetal Growth Retardation Fetus GLUT-3 protein Genes Genetic Genetic Models Glucose Glucose Transporter Grant Growth Growth and Development function Health Homeostasis Hour In Vitro Incidence Induction of Apoptosis Insulin Resistance Knockout Mice Lead Life Light Measurement Metabolic Metabolism Modeling Mus Non-Insulin-Dependent Diabetes Mellitus Nutrient Obesity Organelles Organism Outcome Pathway interactions Patients Pattern Phenotype Physiological Play Pre-implantation Embryo Development Pregnancy Outcome Pregnancy Rate Pregnancy loss Process Production Program Development Proteins Recurrence Recycling Reproductive Medicine Role SLC2A1 gene Staging Techniques Testing Therapeutic Intervention Time Woman Work blastocyst deprivation diabetic embryo/fetus experience glucose transport glucose uptake improved inhibitor/antagonist innovation macromolecule malformation maternal diabetes metabolic abnormality assessment mouse model neuroepithelium novel therapeutics offspring preimplantation prevent programs protein degradation reproductive research study response stressor

项目摘要

DESCRIPTION (provided by applicant): The objective of this proposal is to determine whether autophagy, a highly conserved cellular process, plays a role in mammalian preimplantation development and whether metabolism-induced alterations to this event result in abnormal embryo growth and development. Our mouse models of abnormal embryonic glucose transport, both diabetic and genetic, which we established during the last grant cycle, all experience increased blastocyst apoptosis and result in significant growth deficiencies, some accompanied by specific congenital malformations. Similar growth abnormalities are seen in embryos lacking autophagy proteins as well, and other recent studies suggest that basal autophagy during development is critical to balancing proliferation and apoptosis in response to nutrient supply. Our preliminary data suggest that defects in glucose transport, whether gene knockout mice, antisense mice or controls exposed to abnormal conditions during the first 96 hours post fertilization, persist into adulthood, suggesting that growth deficiencies may be "programmed" in to the embryos from the earliest stages of development. We hypothesize that autophagy may play an important role in this process by regulating the degradation of proteins and also modulating apoptosis. In this case, changes in glucose transport leading to changes in autophagy in the blastocyst stage embryo may program development and pregnancy outcomes. These studies are highly innovative since they are some of the first to examine the role of autophagy in mammalian embryo development and pregnancy outcome. Our rationale is that poor pregnancy outcomes related to fluctuations in glucose availability may be avoidable and/or reversible by changing the pattern of autophagy. These studies aim to test this hypothesis. Specific Aim 1 examines the baseline amount of autophagy in wildtype preimplantation embryos and determines if in vitro manipulation of this process results in blastocyst abnormalities which change pregnancy outcome. Specific Aim 2 evaluates the effect of manipulating autophagy in established mouse models of glucose transporter deficiencies and whether these changes affect embryo development and competence. Specific Aim 3 investigates the possible role GLUT8 may play in autophagy in the mouse blastocyst. The results of this study will establish the role of abnormal protein degradation seen in altered autophagic states during mammalian preimplantation development. Models of excess or deficient autophagy result in embryonic lethality and abnormal neuroepithelium, however, the etiology and timing of this developmental insult are unknown. These proposed experiments will attempt to explore this phenomenon and perhaps provide ideas for therapeutic interventions to improve pregnancy rates in patients with diabetes, recurrent pregnancy loss as well as patients undergoing IVF. PUBLIC HEALTH RELEVANCE: In this proposal, we hypothesize that basal autophagy in the murine blastocyst is regulated by metabolic milieu changes and can alterations in autophagy can result in poor pregnancy outcomes. Blastocysts from diabetic mice exhibit changes in autophagic pathways, as do genetic models of decreased glucose uptake. By modulating the rate of autophagy during the preimplantation period in these models and then transferring these embryos into surrogate mice, the outcomes of these manipulations are determined.

描述（由申请人提供）：本提案的目的是确定自噬（一种高度保守的细胞过程）是否在哺乳动物植入前发育中发挥作用，以及代谢诱导的对该事件的改变是否会导致异常的胚胎生长和发育。我们在上一个资助周期建立的异常胚胎葡萄糖转运小鼠模型，包括糖尿病和遗传性模型，都经历了囊胚凋亡增加并导致显着的生长缺陷，其中一些还伴有特定的先天畸形。在缺乏自噬蛋白的胚胎中也发现了类似的生长异常，最近的其他研究表明，发育过程中的基础自噬对于平衡增殖和细胞凋亡以响应营养供应至关重要。我们的初步数据表明，无论是基因敲除小鼠、反义小鼠还是在受精后 96 小时内暴露于异常条件的对照小鼠，葡萄糖转运缺陷都会持续到成年期，这表明生长缺陷可能从胚胎开始就被“编程”到了胚胎中。最早的发展阶段。我们假设自噬可能通过调节蛋白质降解和调节细胞凋亡在此过程中发挥重要作用。在这种情况下，葡萄糖转运的变化导致囊胚期胚胎自噬的变化，可能会影响发育和妊娠结局。这些研究具有高度创新性，因为它们是首次研究自噬在哺乳动物胚胎发育和妊娠结局中的作用的研究。我们的理由是，通过改变自噬模式，与葡萄糖可用性波动相关的不良妊娠结局可能是可以避免和/或逆转的。这些研究旨在检验这一假设。具体目标 1 检查野生型植入前胚胎中自噬的基线量，并确定该过程的体外操作是否会导致囊胚异常，从而改变妊娠结局。具体目标 2 评估了在已建立的葡萄糖转运蛋白缺陷小鼠模型中操纵自噬的效果，以及这些变化是否影响胚胎发育和能力。具体目标 3 研究 GLUT8 在小鼠囊胚自噬中可能发挥的作用。这项研究的结果将确定在哺乳动物植入前发育过程中自噬状态改变中观察到的异常蛋白质降解的作用。自噬过度或不足的模型会导致胚胎致死和神经上皮异常，然而，这种发育损伤的病因和时间尚不清楚。这些拟议的实验将尝试探索这一现象，并可能为治疗干预提供思路，以提高糖尿病患者、复发性流产以及接受体外受精患者的妊娠率。公共健康相关性：在本提案中，我们假设小鼠囊胚中的基础自噬受到代谢环境变化的调节，并且自噬的改变可能导致不良妊娠结局。糖尿病小鼠的囊胚表现出自噬途径的变化，葡萄糖摄取减少的遗传模型也是如此。通过调节这些模型植入前期间的自噬速率，然后将这些胚胎转移到代孕小鼠中，可以确定这些操作的结果。