Principal Investigator/Program Director (Last, first, middle):
Kleyman, Thomas R.
Pittsburgh Center for Kidney Research
Please complete this form to request use of the Pittsburgh Center for Kidney Research Resources
Principal Investigator:
Institution:
Address:
Phone:
Fax:
Email:
Indicate below the Core resources being requested:
Resources available to Center users:
CORE A: Cellular Physiology
A. Electrophysiologic and functional expression assays of channels and membrane protein in single cell
expression system (Xenopus oocytes, HEK cells)
B. Electrophysiologic and functional expression assays in organized epithelia (voltage clamp measurements of
short-circuit current, TER, capacitance, impedance)
C. Methods of modulating gene expression (recombinant viruses, lipid-mediated transfer, siRNA)
D. Analysis of post-translational modifications of transporters and associated proteins
CORE B: Single Nephron and Metabolomics
A. Microdissection and/or microperfusion of isolated tubules: Measurement of rates of ion transport; Fluorescent
functional (pH, intracellular Ca2+, Na/K/Cl ions) imaging to assess live cell function
B. Single isolated tubule real-time PCR, immunoblotting, immunolabeling and/or enzyme/transporter assays
C. Isolation and analyses of urinary exosomes
D. Renal metabolomics
CORE C: Kidney Imaging
A. Preparation, storage, and analysis of fixed kidney and bladder-associated epithelial cells and tissues,
including imaging capture modalities (brightfield, DIC, darkfield, confocal) and image analysis (stereology)
B. Fine structure analysis, immunoEM localization, scanning electron microscopy, ultrathin cyroelectron
microscopy
C. Real time imaging using spinning disk confocal microscopy, or video microscopy and digital deconvolution,
two-photon microscopy
D. Live animal imaging
CORE D: Model Organisms
A. Yeast expression systems for wild type and disease-causing versions of proteins.
B. Functional in vitro assays using yeast derived factors
C. Establish kidney expressing transgenic zebrafish reporter lines for small molecule genetic screens, targeted
gene expression studies
Training/Mini-Sabbatical request for CORE A __ , CORE B __ , CORE C __ , CORE D __ .
Provide a brief summary of the goal of the project and your proposed studies for which you are requesting
use of the Center resources. Please include a description of the relevance to kidney or urologic diseases,
epithelial cell biology, or physiology:
Principal Investigator/Program Director (Last, first, middle):
Kleyman, Thomas R.
If requesting a mini-sabbatical, describe your goals of the sabbatical:
Indicate source of funding for your planned studies
(please include funding agency, grant number, and PI):
__ Government (NIH, VA, NSF)
__ Private foundation
__ Industry/Corporate
__ Institutional funds
Expenses
The funding provided by the Center applies to services and technical support provided by the Center Cores. In
addition, the Center will provide expert advice and assistance with data analysis, as well as training in relevant
techniques and mini-sabbaticals. Other expenses (such as animals, laboratory supplies, transportation,
accommodations, etc.) are the responsibility of the primary investigator.
Acknowledgment of the Center
For all publications resulting from data obtained using the Center resources, please include the following
acknowledgement: “Studies utilized the NIH sponsored Pittsburgh Center for Kidney Research.”
The Center Director Dr. Thomas Kleyman can provide letters of Center resource availability, consultation or
collaboration upon request.
Please attach, with this request form, a copy of your Biosketch in NIH format including Other Support.