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Histology and Molecular Pathology Tissue arrays (tissue samples from existing archival specimens from various kidney disease models can be provided for exploration and characterization of expression patterns of various novel target molecules) Routine histology Special and/or selective immunohistochemistry RNA scope in situ hybridization Electron microscopic analysis and selected morphometric techniques (e.g., glomerular size, tubulointerstitial fibrosis, GBM width and foot process effacement are provided Laser capture microscopic dissection for special studies of selected renal structures is also offered, with mass spectrometry and/or PCR for molecules of interest in the selected regions Confocal Z section imaging of selected renal structures for detailed spatial analysis can also be performed by the Core In situ mass spectroscopy techniques to optimize spatial resolution of mass spectra in kidney tissues, and propose to fully develop this technique for identification of key molecules in kidney diseases The cell and genome engineering core CRISPR/Cas9 and transposon vectors and expertise in using them for stable genetic modification of kidney cells. Isolate nephron specific primary and immortalized cells from uninjured and injured mice that can either be cultured for future use. In addition, it will isolate protein and/or DNA from cell specific components of injured mouse kidneys for in depth analysis. Isolate nephron specific cells that will be processed for RNA analysis from normal humans and those with renal disease, specifically diabetes. The Phenotyping and Pathophysiology Core (Animal) Phenotyping Subcore Albuminuria Blood urea nitrogen (BUN) Creatinine as serum creatinine or creatinine clearance Glomerular filtration rate (GFR) by FITC inulin Glomerular filtration rate (GFR) by FITC-Sinistrin Plasma renin activity Plasma aldosterone levels Urinary F2-isoprostanes Urinary osmolality Blood and urine Na+, K+ levels Blood pressure by Tail cuff. Blood pressure by Catheterization Blood pressure by Radiotelemetry. Sodium and water balance study with acute salt loading. Necropsy. Kidney Protein and mRNA isolation. Glomerular isolation Injury Model Subcore Kidney transplantation Bone marrow transplantation (BMT) Type II diabetic nephropathy. Type I diabetic nephropathy (e.g. streptozotocin-induced diabetes). 5/6 nephrectomy. Angiotensin II induced nephropathy. Unilateral ureteral obstruction (UUO). Ischemia-reperfusion. Toxic nephropathy. (mercuric chloride, folic acid, Adriamycin, and aristolochic acid induced kidney toxicity) Kidney Oxygen Metabolism and Bioenergetics Subcore Screening and characterization of murine kidney disease model with potential defects in O2 and energy metabolism In vitro and in vivo analysis of partial O2 consumption Training and scientific guidance in the areas of renal hypoxia and metabolism. Quantitation of intermediary metabolism and MALDI imaging mass spectrometry of renal metabolites The In Vivo Mouse Kidney Imaging Subcore BOLD-MRI to investigate renal oxygenation. qBOLD to quantifies local basal oxygen saturation MicroCT: to evaluate three dimensional quantification of renal vascular structure in preclinical models Clinical core General Planning, Implementation, and Evaluation Consultative Service Planning Studios Hypothesis development Study design, Ethics review, Analytic plan and biostatistics Project management services Development of internal and exportable study database, Regulatory support (e.g. IRB, IND, DSMB), Quality assurance/control activities, Data analysis and reporting Biorepository Subcore Access to clinically linked biosamples Blood, urine, DNA, tissue Longitudinal clinical phenotyping in patients with or suspected of kidney disease Histopathologic data Metabolic Phenotyping Unit Subcore (VPMU) Energy expenditure Whole-room indirect calorimeter Metabolic cart systems Portable calorimetry systems Energy and Nutrient intake software Body composition analysis Dual energy x-ray absorptiometry (DXA), Air-displacement plethysmography (BodPod), Bioimpedance Metabolic studies o glucose, amino acid and fatty acid clamp studies o consultation on design and implementation o isotope trace studies for substrate metabolism o consultation on design and implementation Assay measurements o blood, urine and tissue amino acid profiles o blood fatty acid profiles o labeled isotope by MS (leucine, phenylalanine, glucose, palmitic acid) Sodium MRI o skin and muscle tissue Na accumulation measurement Vascular studies o Consultation on Brachial Artery Hyperemia, Pulse Wave Velocity Clinical Epidemiology and Biostatistical Subcore (VCEB) Collaborative interface with large ongoing prospective/retrospective cohorts o Southern Community Cohort Study, Jackson-Heart Study, CARDIA) Consultative services o research question development, design, analytic approach, manuscript/grant evaluation Informatics expertise and consultation o development of clinical decision support, o pharmacogenomics studies, o natural language processing and consultation