Establishment of a mouse model with mutagenesis induced hyperaldosteronism

Spyroglou, Ariadni (2009): Establishment of a mouse model with mutagenesis induced hyperaldosteronism. Dissertation, LMU München: Medizinische Fakultät

Abstract

According to recent epidemiological studies, primary aldosteronism is considered to be responsible for almost 10-15% of all cases of arterial hypertension. The genetic background of this common disease, however, has been elucidated only for the rare familial types whereas in the large majority of sporadic cases genetic causes or modifiers still remain unclear. In an attempt to define novel genetic mechanisms of hyperaldosteronism we utilized a random mutagenesis screen after treatment with the alkylating agent N-ethylnitrosourea (ENU) and phenotypically characterized affected mice for their blood aldosterone levels. As the detection method we used a time resolved fluorescent immunoassay which allows the measurement of aldosterone in very small murine plasma volumes. Using this assay we determined the normal aldosterone values for C3HeB/FeJ wild type mice under baseline conditions and following specific stimulation and suppression tests. We could demonstrate the expected increase in aldosterone response upon ACTH stimulation, a high potassium diet and an angiotensin II injection, as well as the decrease after a dexamethasone suppression test, a normal saline load test and a fludrocortisone suppression test. On the molecular level, the expression of aldosterone synthase showed a similar pattern with a fast response to the investigated stimuli. These tests should later be applied to the mouse lines derived from the ENU screen, in order to investigate potential abnormal response to these stimuli in comparison to wild type animals. Furthermore, aldosterone measurement was carried out in more than 2000 F1 offspring (of both genders) of chemically mutated inbred C3HeB/FeJ mice. From these tested F1 offspring, aldosterone levels were consistently elevated (defined as levels above 3 SD over the mean of untreated animals)upon repeated measurement in 9 animals (8 females and 1 male). Further breeding of affected female animals gave rise to F2 pedigrees from which four established lines displayed high aldosterone values. These animals served for a detailed phenotypic characterization and showed an increased aldosterone to renin ratio, low potassium values and normal renal function in line with the presence of primary aldosteronism. In addition, the investigation of their cardiac phenotype showed increased collagen deposits and subsequently cardiac fibrosis, as also observed in patients suffering from primary aldosteronism. In the future, genetic SNP analysis can be performed to identify underlying genetic loci, responsible for this trait. Taken together, these data demonstrate the feasibility of a phenotype-driven mutagenesis screen to detect and establish mutant mouse lines with a high aldosterone phenotype.

• Dokumententyp: Dissertationen (Dissertation, LMU München)
• Themengebiete: 600 Technik, Medizin, angewandte Wissenschaften ; 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
• Fakultäten: Medizinische Fakultät
• Sprache der Hochschulschrift: Englisch
• Datum der mündlichen Prüfung: 18. Juni 2009
• 1. Bericht­erstatter:in: Beuschlein, Felix