Microfluidic Platform for Probing Ceramide Channels

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DESCRIPTION (provided by applicant): A microfluidic platform is proposed that will aid in investigating processes involved in lipid channel formation and dissolution, leading to a deeper understanding of molecular transport across cell membranes, and ultimately contribute to our knowledge of biological channels and their relationships to disease processes. The project will leverage recent results from our team, including the first demonstration of single-molecule detection within a microfluidic ion channel chip using a single membrane-bound biological ion channel as a conductometric sensing element, with real-time control of analyte concentration and buffer conditions on either side of the channel. In this proposal, the proof-of-concept microfluidic platform will be extended to demonstrate in situ phospholipid membrane formation, integrated supporting gels and microporous films for enhanced membrane stability, automated multiplexed lipid channel measurements, and thin film thermal control at the membranes to enable the observation of the relationships between enthalpy and membrane formation, channel formation and dynamics, and analyte/channel interactions. A range of experiments that are not feasible or extremely time-consuming with existing technology will be conducted, with a focus on investigating channels formed by ceramide, a sphingolipid implicated in apoptosis. The structure and function of ceramide channels will be probed using the microfluidic system, with controlling factors manipulated to investigate their impacts on the regulation of channel size and stability. This effort will result in a novel and unique electrophysiology platform which may be applied to a wide range of fundamental and applied investigations of biological channels, together with experimental results that will expand our understanding of an important lipid channel system involved in programmed cell death. PUBLIC HEALTH RELEVANCE: A unique experimental platform is proposed that will elucidate the processes involved inbiological membrane channel formation and dissolution, leading to a deeper understanding of molecular transport across cell membranes, and ultimately contribute to our knowledge of biological channels and their relationships to disease processes.