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High‐Throughput Production and Phenotyping of Rat Knockout Models for Hypertension
Author(s) -
Moreno Carol,
Dwinell Melinda R,
Majewski Rebecca R,
Kwitek Anne E,
Jessner Martin J,
Feroah Thom R,
Forster Hubert V,
Mattson David L,
Lombard Julian H,
Macquestion Gregory D,
Roman Richard J,
Greene Andrew S,
Cowley Allen W,
Jacob Howard J
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1236
Subject(s) - gene , biology , medicine , tilling , endocrinology , mutant , mutagenesis , blood pressure , wild type , candidate gene , allele , knockout mouse , genetics
The inability to generate knock‐out (KO) rats has been a major limitation for studying gene function in rats. PhysGen‐MCWuses a strategy that combines ENU mutagenesis to induce spermatic gene mutations, with TILLING, a high‐throughput assay which identifies mutations between wild‐type and mutated alleles to confirm and identify KO rats. The goal is to identify the induced point mutations in 112 selected genes in three different rat strains Dahl Salt‐Sensitive, Brown Norway, and Fawn‐Hooded Hypertensive rat with the broad goal of developing 99 total strains of KO rats (33 genes on 3 different genome backgrounds), which are then characterized physiologically for of cardiovascular, pulmonary and behavioral traits. We have currently developed 26 gene‐mutation strains, of which six are KO for different genes. Males are injected with a variable dose of ENU and then bred to wild‐type females to produce the KO rats, which is then brought to homozygosis. An ultrasound protocol assesses cardiac function by echocardiography in basal and stressed (dobutamine administration) conditions. Vascular function is studied in aortic rings with responsiveness to vasoconstrictors and vasodilators. Plasma levels 29 hormones, electrolytes and blood indicators are also measured. Blood pressure is determined and kidney function is assessed by protein excretion. KO rats availability and data is posted in the PGA web site periodically (pga.mcw.edu).