Animal Model, Analytical Chemistry & Biochemistry
Core Team: Steve Kinsey & Michelle Izuka
This multifunction core provides support for the PPG in the following areas:
- Generation of an experimental model of hypertension in the rat. The model of choice is the ouabain hypertensive rat. This induced form of hypertension and its matched controls are the source for various preparations including intact arteries, vascular cells and blood.
- Measurement of endogenous and exogenous ouabain (and other compounds) in the circulation of animals.
- Breeding and genotyping of transgenic mice.
- Preparation of acutely dissociated myocytes from rat and mice arterioles.
The Core provides new and/or unique tools for extensive use by the various projects of the PPG. The Core provides widely used preparations on a routine basis that would otherwise prove to be burdensome and/or beyond the expertise of the individual projects of the PPG. The Core also coordinates the PPG projects so that preparations can be studied (when appropriate) simultaneously by different methods and laboratories.
Specific Core Functions
Ouabain Hypertensive Rats - Generation of the model and controls, documenting blood pressures, measuring key circulating hormones, and providing vascular tissue from rats at defined time points from the prehypertensive state through to chronic hypertension (Manunta et al., 1994).
Assay of Endogenous Ouabain and Ouabain in Rats and Mice - Both immunoassay (Manunta et al., 1994) and mass spectrometric methods (see below) are employed to document the circulating (and tissue) levels as needed.
LC-MS–MS of Endogenous Ouabain in Normal Rat Plasma
- Transgenic Mice - The core supervises the breeding of various lines of transgenic mice (with altered Na+ pump α1 or α2 subunits or Na/Ca exchanger – type 1) and ensures that sufficient mice are available to the projects as needed.
- Validation of Transgenic Animals - All transgenic mice are monitored for unambiguous proof of genotype. Identified animals are selected for experimental use and/or breeding.
- Preparation of Vascular Myocytes - The core will prepare freshly isolated arterial smooth muscle cells on a routine basis.
Manunta P, Rogowski AC, Hamilton BP, Hamlyn JM. Ouabain-induced hypertension in the rat: relationships among plasma and tissue ouabain and blood pressure. J. Hypertens. 12:549-560, 1994.