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+）是细胞中第二丰富的阳离子。由于Mg 2+在数百种酶促反应中作为辅因子，它对许多生物活性具有深远的影响，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+缺乏与各种疾病状态如动脉粥样硬化、高血压、糖尿病、哮喘和酒精中毒有关并不奇怪。然而，我们对Mg2+稳态的理解远远落后于其他阳离子，如Ca2+，Na+，K+和H+，主要是因为Mg2+的转运蛋白尚未在分子水平上确定。此外，很少有工作已经进行了关于镁离子缺乏对基因表达的影响，特别是关于其在各种病理条件下的作用。使用cDNA微阵列进行基因表达谱分析是鉴定参与特定细胞或疾病过程的基因的有力方法。我们建议使用这项技术来确定参与镁稳态的基因，并阐明镁缺乏导致各种疾病状态的机制。通过比较正常小鼠与缺镁小鼠的选择器官的全局基因表达模式，应该可以鉴定参与镁稳态的基因，例如编码镁转运蛋白的基因。在缺镁条件下异常表达的基因也应该被鉴定出来。这将有助于更好地了解镁在健康和疾病中的作用。在目标1和2中，商业cDNA微阵列将被用于研究急性和慢性镁缺乏对基因表达的影响。目的3：从镁缺乏和镁充足小鼠的不同器官中构建抑制消减杂交文库。然后将来自文库的克隆用于构建用于基因表达谱分析的定制cDNA微阵列。这种技术的组合应导致鉴定在缺镁期间差异表达的所有基因。