Pittsburgh Center for Kidney Research
Summary
The O’Brien Kidney Research Center at the University of Pittsburgh and Icahn School of Medicine at Mount Sinai seeks to enhance the efficiency and productivity of a large number of kidney-related research projects currently in progress; facilitate acquisition of data for new projects; promote collaborations among center investigators; and provide tools to facilitate translational research. These functions depend on the Center's Biomedical Cores: Physiology Core, Model Systems Core, and KIDNIT (Kidney Imaging) Resource Development Core. Our Cores provide physiological, cell biological, genetic, analytical, molecular biological, and drug discovery tools, as well as model organisms.
Physiology Core
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Description
The overall goal of the Physiology Core is to elucidate, at a molecular and cellular level, the function and regulation of key proteins involved in kidney health and disease. The Core provides in vitro and in vivo technologies to explore the function and regulation of membrane transporters, channels, and other resident proteins using single molecule approaches, model organisms, and native epithelia. Services provided are listed below:
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Services
- Data analysis - cellular and whole animal physiological studies, or imaging studies
- Epithelial transport in Ussing chambers
- Functional imaging analyses
- In vitro microperfusion of isolated tubules for functional fluorescence assays of single cell function
- In vitro microperfusion of isolated tubules for measurement of transepithelial ion and solute transport
- Isolation of single nephron segments for immunofluorescence, enzyme assays, RNA and protein isolation
- Isolation of Xenopus laevis oocytes
- Manuscript development
- Nephron segment microdissection/isolation and microperfusion training
- Oocyte two-electrode voltage clamp
- Patch-clamp analyses
- Patch-clamp training
- Quantification of surface protein expression
- Training on use of microinjectors
- Training on use of micropipette pullers and microforges to craft micropipettes for patch-clamp and in vitro microperfusion
- Two-electrode voltage clamp training
- Xenopus laevis technologies training
Model Systems Core
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Description
The overall goal of the Model Systems Core is to provide a diverse array of innovative model systems ranging from yeast, tocultured kidney cells and organoids, to whole animal rodent models to understand and explore kidney development, function, and systemic physiology in normal and disease states. Core directors will facilitate integration between these model systems, enabling investigators to test hypotheses at multiple resolutions from molecules to cells to tubules to whole animals. Services provided are listed below:
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Services
- Animal Handling and Husbandry
- Animal Models - Blood pressure telemetry
- Animal Models - Mitochondrial / Bioenergetics (Seahorse)
- Animal Models - Purine metabolomics
- Animal Models - Injury biomarker assessment
- Animal Models - Measurement of GFR and RBF
- Animal Models - Metabolic cage studies, urine metabolites
- Animal Models - Real-time super-resolution ultrasound of kidney microvasculature
- Animal Models - Tissue Harvesting
- Data analysis - cellular and whole animal physiological studies, or imaging studies
- Disease Models: AKI (cecal ligation and puncture)
- Disease Models: AKI (cisplatin nephrotoxicity)
- Disease Models: AKI (gentamicin nephrotoxicity)
- Disease Models: AKI (ischemia-reperfusion injury)
- Disease Models: AKI (sickle cell crisis)
- Disease Models: CKD (aristolochic acid)
- Disease Models: CKD (chronic angiotensin II infusion)
- Disease Models: CKD (FSGS)
- Disease Models: CKD (genetic models)
- Disease Models: CKD (two kidney one clip hypertension)
- Disease Models: CKD (unilateral ureteral obstruction)
- Epithelial Models - Mammalian cell culture models (proximal tubule, collecting duct, polarized epithelia)
- Epithelial Models - Organoid models to study kidney injury
- Epithelial Models - Organoid models to study kidney physiology
- Epithelial Models - Organoid models to study nephron development
- HPLC-Fl - ATP Metabolome
- Manuscript development
- Purine Metabolome by UPLC-MS/MS
- Special Rodent Diets (low and high Na+ and K+)
- Yeast Models - Positive genetic selection assays
- Yeast Models - Protein quality control (degradation, ubiquitylation) assays
- Yeast Models - Bioinformatic analysis of genotype-phenotype correlations (TopMed, Clinvar, UK BioBank)
- Yeast Models - Computational identification of functional variants
- Yeast Models - Protein trafficking assays (immunofluorescence, density centrifugation)
Kidney Imaging: Developing Novel and Innovative Tools (KIDNIT) Development Core
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Description
The KIDNIT Resource Development Core provides a national resource for investigators who require detailed and quantitative morphological analysis of kidney-associated cells and tissues, particularly those that are epithelial in nature, as well as whole organs. Services include:
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Services
- Confocal microscopy - Laser scanning confocal microscopy
- Confocal microscopy - Spinning disk confocal microscopy
- Confocal microscropy - Fluorescent lifetime imaging
- Confocal microscropy - Fluorescent recovery after photobleaching
- Confocal microscropy - Ribbon scanning confocal microscopy for cleared organs/tissues
- Consultation for development of new approaches to study the kidney and lower urinary tract using morphological tools
- Data analysis - cellular and whole animal physiological studies, or imaging studies
- Electron Microscopy - CLEM
- Electron Microscopy - Freeze fracture microscopy
- Electron Microscopy - Freeze substitution
- Electron Microscopy - Immuno-Electron microscopy
- Electron Microscopy - Negative staining
- Electron Microscopy - Platinum replica electron microscopy
- Electron Microscopy - Scanning electron microscopy (field-emission)
- Electron Microscopy - Transmission electron microscopy
- Electron Microscopy - Ultramicrotomy
- Electron Microscopy - Ultrathin cryo EM
- Image Analysis - Imaris
- Image Analysis - Leica LASX
- Image Analysis - Metamorph
- Image Analysis - NIH image/FIJI
- Image Analysis - NIS elements
- Image Analysis - Photoshop
- iPOL imaging
- Live-cell microscopy - Fluorescent lifetime imaging
- Live-cell microscopy - Fluorescent recovery after photobleaching
- Live-cell microscopy - Incubator microscopy
- Live-cell microscopy - Lattice light sheet microscopy
- Live-cell microscopy - Spinning disk confocal microscopy
- Live-cell microscopy -Microinjection
- Manuscript development
- Meso-Spim light sheet microscopy
- Multiphoton microscopy - Fluorescent lifetime imaging
- Multiphoton microscopy - Fluorescent recovery after photobleaching
- Multiphoton microscopy - Intravital imaging
- Multiphoton microscopy - Multiphoton confocal microscopy
- Standard light microscopy - Brightfield, darkfield, epifluorescence, DIC
- Standard light microscopy - Macro (stereo) imaging microscopy
- Super-resolution microscopy - AX/NSPARC
- Super-resolution microscopy - CREST
- Super-resolution microscopy - STED
- Super-resolution microscopy - STORM
- Training in live-cell and intravital approaches to study the biology/pathobiology of kidney and bladder tissues
- Training in quantitative image analysis
- Training in sample preparation of kidney and bladder tissues for light and electron microscopic analysis
- Training in specialized sample preparation to perform labeling and analysis of cleared organs and tissues
- TRIF Microscopy - TIRF imaging acquisition and analysis
- Whole slide scanning - brightfield and epifluorescence
Contact Information
If you would like more information regarding services provided by Pittsburgh Center for Kidney Research, please contact:
Thomas Kleyman
Ora Weisz
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