Publication Date

2011

Document Type

Thesis

Committee Members

Thomas Brown (Advisor), Courtney Sulentic (Committee Member), Christopher Wyatt (Committee Member)

Degree Name

Master of Science (MS)

Abstract

AMP-activated protein kinase (AMPK) regulates cellular metabolism and promotes ATP production when energy is depleted. Evidence suggests that AMPK may be involved in oxygen sensing by carotid body cells, which are responsible for regulating the breathing rate to maintain proper blood oxygen levels. There are two isoforms of the catalytic alpha subunit, AMPKalpha1 and AMPKalpha2, which could be involved in oxygen sensing. Here, the production of a short hairpin RNA (shRNA) targeting both catalytic isoforms of AMPK in human, mouse, and rat is described. The shRNA causes significant knockdown of both isoforms of AMPK alpha in mouse and human cells and a significant reduction in AMPK activity, measured as phosphorylation of a direct target. This shRNA will be used to generate a rat model with tissue-specific knockdown of AMPKalpha1 and 2 using a Cre-Lox recombination system to determine the role of AMPK in oxygen sensing by the carotid body.

Page Count

72

Department or Program

Microbiology and Immunology

Year Degree Awarded

2011


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