SCHOOL OF MEDICINE
Expression of GFP in the renal tubules of a living rat following subcapsular injection of an AAV9-GFP (Xiao Xiao, Ken Dunn and George Rhodes)
Expression of GFP in the pancreas of a living rat following subcapsular injection of an AAV8-GFP (Xiao Xiao, Ken Dunn and Jenn Ryan)
Time series of images collected from the liver of a living mouse whose white cells were labeled in vivo with pH-sensitive liposomes containing calcium (Rudy Juliano, Takashi Hato and Pierre Dagher)
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 IU 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 IU 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.
We apply, refine and extend 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. We develop novel rat models providing titratable expression of introduced genes and transgenic rats to support novel in vivo studies of the rat glomerulus.
Protocol Delivery of Fluorescent Probes to the Kidney.
In-house vector constructs information and Sequence.
Robert L. Bacallao, MD
Richard Day, PhD
☎: (317) 274-2166
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