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Sepsis is a leading cause of acute kidney injury (AKI) and mortality in children. In order to develop new therapies, it is critical to understand the development of pediatric sepsis and its effects...

Sepsis is caused by a severe microbial infection and can lead to acute kidney injury (AKI). This results in a high mortality rate of 70%. No specific therapies exist for septic AKI. To test if mito...

Corticotropin releasing hormone receptor CRHR and the VT2 arginine vasotocin receptor VT2R are vital links in the hypothalamic-pituitary-adrenal axis that enable a biological response to stressful stimuli in avian species. CRHR and VT2R are both G-protein coupled receptors GPCRs, and have been shown by us to form a heterodimer via fluorescent resonance energy transfer FRET analysis in the presence of their respective ligands, corticotrophin re- leasing hormone CRH and arginine vasotocin AVT. The dimeriza- tion interface of the heterodimer is unknown, but computational analyses predict transmembrane domains TMs as likely sites of the interaction. We constructed chimerical VT2Rs, tagged at the C-terminal ends with either cyan fluorescent protein CFP or yellow fluorescent protein YFP, by replacing the fourth transmembrane re- gion TM4 of VT2R with TM4 of the 2-adrenergic receptor 2AR. The VT2R/2AR chimeras were expressed in HeLa cells and proper trafficking is confirmed by observing cell membrane localization us- ing confocal microscopy. VT2R/2AR-YFP chimera functionality was confirmed with a Fura-2 acetoxymethyl ester Fura-2AM assay. FRET analysis was then performed on VT2/2AR-chimera/CRHR pairs, and the calculated distance was observed to be 10 nm apart, indicating that heterodimerization was partly disrupted by mutating TM4 of the VT2R. Therefore, TM4 may form one region of the possible dimeriza- tion interfaces between the VT2R and CRHR. © 2008 Society of Photo-Optical