Genetically encoded biosensor proteins enable the detection of the spatial and temporal characteristics of specific cell signaling or metabolic events in single living cells. When used in combination with intravital microscopy (IVM), the changing signals from the biosensor probes can be monitored from the tissues of living animals, supporting unique studies of cell biology in the relevant physiological context of the living animal. During the previous grant cycle the O’Brien Center Probe Development Core developed novel fluorescent biosensor reagents and protocols for imaging these probes in vivo using multiphoton microscopy. In parallel, the O’Brien Center Probe Delivery Core developed methods of in vivo gene transfer to provide investigators with a flexible approach for utilizing fluorescent protein biosensors, obviating the time and costs associated with generation of transgenic animals.
The core applies, refines and extends the approaches developed during the previous grant period to provide renal investigators with effective new reagents and protocols to support unique studies of renal cell biology in vivo. The Biosensor Development and Delivery Core develops several important resources for the renal research community. We provide optimized biosensor probes that have been validated for IVM, provide these probes in viral vectors optimized for in vivo gene transfer and provide protocols for delivery and use of probes. The core develops novel rat models providing titratable expression of introduced genes and transgenic rats to support novel in vivo studies of the rat glomerulus.