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University Pittsburgh 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

  • 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:

  • Services
    • Available Resources
      • Isolation of single nephron segments for immunofluorescence, enzyme assays, RNA and protein isolation
      • In vitro microperfusion of isolated tubules for measurement of transepithelial ion and solute transport
      • In vitro microperfusion of isolated tubules for functional fluorescence assays of single cell function
      • Isolation of Xenopus laevis oocytes
      • Oocyte two-electrode voltage clamp
      • Quantification of surface protein expression
      • Patch-clamp analyses
      • Epithelial transport in Ussing chambers
      • Functional imaging analyses
      • Other procedures
    • Hands-on Training
      • Xenopus laevis technologies
      • use of microinjectors
      • use of micropipette pullers and microforges to craft micropipettes for patch-clamp and in vitro microperfusion
      • Two-electrode voltage clamp
      • Ussing chamber systems
        • Patch-clamp
        • Nephron segment microdissection/isolation and microperfusion

Model Systems Core

  • 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:

  • Services
    • Yeast Models Subcore
      • Protein quality control (degradation, ubiquitylation) assays
      • Protein trafficking assays (immunofluorescence, density centrifugation
      • Positive genetic selection assays
      • Computational identification of functional variants
      • Bioinformatic analysis of genotype-phenotype correlations (TopMed, Clinvar, UK BioBank)
    • Epithelial Models Subcore
      • Mammalian cell culture models (proximal tubule, collecting duct, polarized epithelia)
      • Organoid models to study nephron development
      • Organoid models to study kidney physiology
      • Organoid models to study kidney injury
    • Animal Models Subcore
      • Measurement of GFR and RBF
      • Metabolic cage studies, urine metabolites
      • Blood pressure telemetry
      • Cardiac Function (pressure-time parameters)
      • Real-time super-resolution ultrasound of kidney microvasculature
      • Purine metabolomics
      • Mitochondrial / Bioenergetics (Seahorse)
      • Injury biomarker assessment
      • Disease Models: AKI (ischemia-reperfusion injury)
      • Disease Models: AKI (cisplatin nephrotoxicity)
      • Disease Models: AKI (gentamicin nephrotoxicity)
      • Disease Models: AKI (cecal ligation and puncture)
      • Disease Models: AKI (sickle cell crisis)
      • Disease Models: CKD (genetic models)
      • Disease Models: CKD (FSGS)
      • Disease Models: CKD (unilateral ureteral obstruction)
      • Disease Models: CKD (two kidney one clip hypertension)
      • Disease Models: CKD (chronic angiotensin II infusion)
      • Special Rodent Diets (low and high Na+ and K+)

Kidney Imaging: Developing Novel and Innovative Tools (KIDNIT) Development Core

  • 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:

  • Services
    • Consultation and Training opportunities
      • Consultation for development of new approaches to study the kidney and lower urinary tract using morphological tools.
      • 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.
      • Training in quantitative image analysis.
      • Training in live-cell and intravital approaches to study the biology/pathobiology of kidney and bladder tissues.
    • Standard light microscopy
      • Brightfield, darkfield, epifluorescence, DIC
      • Macro (stereo) imaging microscopy
    • Confocal microscopy
      • Laser scanning confocal microscopy
      • Spinning disk confocal microscopy
      • Ribbon scanning confocal microscopy for cleared organs/tissues
      • Fluorescent lifetime imaging
      • Fluorescent recovery after photobleaching
    • Multiphoton microscopy
      • Multiphoton confocal microscopy
      • Fluorescent lifetime imaging
      • Fluorescent recovery after photobleaching
      • Intravital imaging
    • TIRF microscopy
      • TIRF imaging acquisition and analysis
    • Super-resolution microscopy
      • STED
      • CREST
      • STORM
      • AX/NSPARC
    • Live-cell microscopy
      • Spinning disk confocal microscopy
      • Fluorescent lifetime imaging
      • Fluorescent recovery after photobleaching
      • Lattice light sheet microscopy
      • Incubator microscopy
      • Microinjection
    • Large Volume Light-Sheet microscopy
      • Meso-Spim light sheet microscopy
    • Additional imaging modalities
      • Whole slide scanning - brightfield and epifluorescence
      • iPOL imaging
    • Electron microscopy
      • Scanning electron microscopy (field-emission)
      • Transmission electron microscopy
      • Freeze fracture microscopy
      • Platinum replica electron microscopy
      • Negative staining
      • Freeze substitution
      • Ultramicrotomy
      • Ultrathin cryo EM
      • CLEM
      • Immuno-Electron microscopy
    • Image analysis
      • Photoshop
      • Imaris
      • Metamorph
      • NIS elements
      • Leica LASX
      • NIH image/FIJI

Contact Information

If you would like more information regarding services provided by Pittsburgh Center for Kidney Research, please contact:

Thomas Kleyman This email address is being protected from spambots. You need JavaScript enabled to view it. or
Ora Weisz This email address is being protected from spambots. You need JavaScript enabled to view it.
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