The primary aim of the proposed work is to develop microelectrodes for determining ifnasal membrane cholesterol is elevated in human cystic fibrosis. Studies will calibratethe electrodes through solution experiments and validate this calibration using a newproposed influx titration assay where the electrode is used as an indicator of influx.Human studies using the electrodes will involve single cell studies on epithelial cellsfrom nasal scrapes. In vivo electrode measurements in the human nose are alsoproposed. In developing the measurements as a diagnostic tool, implementation of invitro measurements at cells and mouse tissues is proposed to establish the relationshipbetween CFTR function and plasma membrane cholesterol content.
Microelectrodes are used to measurer cholesterol efflux from cells andtissues. In vivo experiments are proposed in the human nose to test thehypothesis that cholesterol efflux rate can be used as a diagnostic tool in thestudy of human cystic fibrosis.
