A Study of a Proposed Molecular Mechanism for Idiopathic Cases of Hypertension: Detection of Aldosterone-receptor Complex at Period Homolog 1 (Per1) Promoter Using Chromatin Immunoprecipitation (ChIP)
Bachelor of Science
Major or Concentration
Hypertension severely affects the quality and quantity of life of those afflicted with the disease. In most cases where the cause is known, hypertension is due to an abnormally high sodium condition; however, the mechanism by which this occurs is unknown. This phenomenon is particularly detrimental to patients considered to be non-dippers, experiencing no nocturnal dip in blood pressure. A proposed molecular mechanism for the relationship between dipping and sodium concentration involves the regulation of the circadian gene Period 1 homolog (Per1) by aldosterone and was studied by using chromatin immunoprecipitation (ChIP). This technique results in an enrichment of the chromatin fragments that are bound by the protein of interest, which after Polymerase Chain Reaction (PCR), it can be verified whether or not aldosterone directly increases the expression of Per1. Primers were successfully designed for two locations within the Per1 gene. One set was for the -156 glucocorticoid regulatory element (GRE), a potential region where the aldosterone receptor binds and a second set to a 3’ was designed as a control. Optimization of the enzymatic shearing conditions was determined to be ten minutes. There were no PCR products for the samples after the complete ChIP assay; however, the expected results were obtained for those samples without pre-clearing of chromatin and antibody treatment. For future study, samples should be tested systematically using PCR to determine in which step the DNA fragments are lost.
Turner, Brent Colin, "A Study of a Proposed Molecular Mechanism for Idiopathic Cases of Hypertension: Detection of Aldosterone-receptor Complex at Period Homolog 1 (Per1) Promoter Using Chromatin Immunoprecipitation (ChIP)" (2009). Student Research Submissions. 2.