The Biochemical Core Laboratory of this Program Project Grant will provide a consolidated highly specialized well-equipped and professional unit capable of performing a variety of radioimmunoassay, biochemical and molecular procedures as required by project investigators. No single investigator laboratory supported by R01 would be capable of equipping and staffing this resource. The Core will carry out all the analytical techniques: 1) Assay of CYP and NOS activities by radiochemical HPLC. 2) Measurement of 20- HETE and EETs in tissue samples, CSF and microdialysates by LC/MS. 3) Identification of novel metabolites generated by CYP4F and CYP4X enzymes by LC?MS/MS. This Core is also equipped to measure IP3, DAG, microglial and astroglial products in the brain and cerebral vasculature. In addition we will measure CYP and NOS protein using western blots and immunohistochemistry. Molecular measurements with real time RT-PCR and in situ hybridizations will be carried out as outlined in the projects. Access to the analytical expertise and equipment in the central facility is essential to each of the proposed projects and justified by the large number of biochemical and molecular determinations required for successful completion of the proposed studies.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL059996-08
Application #
7342767
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
2007-02-01
Project End
2010-01-31
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
8
Fiscal Year
2007
Total Cost
$304,528
Indirect Cost
Name
Medical College of Wisconsin
Department
Type
DUNS #
937639060
City
Milwaukee
State
WI
Country
United States
Zip Code
53226
Gebremedhin, Debebe; Zhang, David X; Carver, Koryn A et al. (2016) Expression of CYP 4A ?-hydroxylase and formation of 20-hydroxyeicosatetreanoic acid (20-HETE) in cultured rat brain astrocytes. Prostaglandins Other Lipid Mediat 124:16-26
Pabbidi, Mallikarjuna R; Mazur, Olga; Fan, Fan et al. (2014) Enhanced large conductance K+ channel activity contributes to the impaired myogenic response in the cerebral vasculature of Fawn Hooded Hypertensive rats. Am J Physiol Heart Circ Physiol 306:H989-H1000
Gebremedhin, Debebe; Gopalakrishnan, Sandeep; Harder, David R (2014) Endogenous events modulating myogenic regulation of cerebrovascular function. Curr Vasc Pharmacol 12:810-7
Carver, Koryn A; Lourim, David; Tryba, Andrew K et al. (2014) Rhythmic expression of cytochrome P450 epoxygenases CYP4x1 and CYP2c11 in the rat brain and vasculature. Am J Physiol Cell Physiol 307:C989-98
Gebremedhin, Debebe; Terashvili, Maia; Wickramasekera, Nadi et al. (2013) Redox signaling via oxidative inactivation of PTEN modulates pressure-dependent myogenic tone in rat middle cerebral arteries. PLoS One 8:e68498
Wickramasekera, Nadi T; Gebremedhin, Debebe; Carver, Koryn A et al. (2013) Role of dual-specificity protein phosphatase-5 in modulating the myogenic response in rat cerebral arteries. J Appl Physiol 114:252-61
Yang, Zeng-Jin; Carter, Erin L; Kibler, Kathleen K et al. (2012) Attenuation of neonatal ischemic brain damage using a 20-HETE synthesis inhibitor. J Neurochem 121:168-79
Renic, Marija; Kumar, Suresh N; Gebremedhin, Debebe et al. (2012) Protective effect of 20-HETE inhibition in a model of oxygen-glucose deprivation in hippocampal slice cultures. Am J Physiol Heart Circ Physiol 302:H1285-93
Liu, Xiaoguang; Li, Chunyuan; Falck, John R et al. (2012) Relative contribution of cyclooxygenases, epoxyeicosatrienoic acids, and pH to the cerebral blood flow response to vibrissal stimulation. Am J Physiol Heart Circ Physiol 302:H1075-85
Terashvili, M; Sarkar, P; Nostrand, M V et al. (2012) The protective effect of astrocyte-derived 14,15-epoxyeicosatrienoic acid on hydrogen peroxide-induced cell injury in astrocyte-dopaminergic neuronal cell line co-culture. Neuroscience 223:68-76

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