Magnesium Deficiency: Global Gene Expression Study

镁缺乏症：全球基因表达研究

• 批准号: 6503795
• 负责人: PATRICK John SCHULTHEIS
• 金额: $ 12.98万
• 依托单位: NORTHERN KENTUCKY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6503795
• 关键词: complementary DNA; gene expression; genetic library; genetic models; homeostasis; ion transport; laboratory mouse; magnesium deficiency; magnesium ion; malnutrition; microarray technology; molecular cloning; nutrition related tag; subtraction hybridization; transport proteins

DESCRIPTION (provided by applicant): Magnesium (Mg2+) is the second most abundant cation in cells. Because Mg2+ serves as a cofactor in hundreds of enzymatic reactions, it has a profound influence on many biological activities including DNA and protein synthesis, ion transport, intracellular signal transduction, and cell growth and differentiation. Consequently, it is not surprising that Mg2+ deficiency has been implicated in various disease states such as atherosclerosis, hypertension, diabetes, asthma, and alcoholism. Yet, our understanding of Mg2+ homeostasis lags far behind that of other cations such as Ca2+, Na+, K+, and H+, largely because the transporters of Mg2+ have not been identified at the molecular level. In addition, little work has been conducted concerning the effects of Mg2+ deficiency on gene expression, especially with respect to its role in various pathological conditions. Gene expression profiling using cDNA microarrays represents a powerful method for identifying genes involved in specific cellular or disease processes. We propose to use this technology to identify genes involved in magnesium homeostasis and to elucidate the mechanisms by which magnesium deficiency leads to various disease states. By comparing global gene expression patterns of select organs of normal mice to their counterparts in magnesium-deficient mice, it should be possible to identify genes involved in magnesium homeostasis, such as those encoding magnesium transporters. Genes that are aberrantly expressed under magnesium-deficient conditions should also be identified. This should lead to a better understanding of magnesium's role in health and disease. In Aims 1 and 2 commercial cDNA microarrays will be used to investigate the effects of acute and chronic magnesium deficiency on gene expression, respectively. In Aim 3 suppression subtraction hybridization libraries will be constructed from various organs of magnesium-replete and magnesium-deficient mice. Clones from the library will then be used to construct custom cDNA microarrays for gene expression profiling. This combination of techniques should lead to the identification of all genes that are differentially expressed during magnesium deficiency.

描述（由申请人提供）：镁（Mg2+）是细胞中第二大阳离子。由于MG2+在数百种酶促反应中充当辅因子，因此它对许多生物学活性都有深远的影响，包括DNA和蛋白质合成，离子转运，细胞内信号转导以及细胞生长和分化。因此，MG2+缺乏症已与各种疾病状态有关，例如动脉粥样硬化，高血压，糖尿病，哮喘和酒精中毒也就不足为奇了。然而，我们对MG2+稳态的理解落后于CA2+，Na+，K+和H+等其他阳离子的理解，这很大程度上是因为在分子水平上尚未确定MG2+的转运蛋白。此外，关于MG2+缺乏对基因表达的影响，尤其是在各种病理条件下的作用方面，几乎没有进行任何工作。使用cDNA微阵列进行基因表达分析是一种鉴定与特定细胞或疾病过程有关的基因的强大方法。我们建议使用该技术来识别镁稳态涉及的基因，并阐明镁缺乏症导致各种疾病状态的机制。通过将正常小鼠的精选器官的全局基因表达模式与镁缺陷小鼠的对应物进行比较，应该可以鉴定涉及镁稳态的基因，例如编码镁转运蛋白的基因。还应鉴定出在镁缺陷条件下异常表达的基因。这应该可以更好地了解镁在健康和疾病中的作用。在AIMS 1和2中，商业cDNA微阵列将分别用于研究急性和慢性镁缺乏对基因表达的影响。在AIM 3抑制减法杂交文库中，将由镁的各种器官和缺乏镁的小鼠构建。然后，图书馆的克隆将用于构建用于基因表达分析的自定义cDNA微阵列。这种技术的组合应导致鉴定在镁缺乏期间差异表达的所有基因。