In response to fluctuations in environmental oxygen in the cells of the blood brain barrier (BBB) undergo a number of complex adaptive measures in order to maintain tissue homeostasis and hemostasis. These adaptive responses are particularly important when the balance of oxygen availability and utilization is altered as a result of the pathophysiology of CNS disease. At the microvascular level oxygen responsive signaling mechanisms involving both the endothelial cell and the pericyte regulate angiogenesis, vascular permeability and metabolism. We propose to investigate early responses of the CNS pericyte following in vitro exposure to hypoxia using the GaSPak 100 hypoxia system (Becton Dickinson and Company Sparks, MD.) Within 15 minutes of exposure to low oxygen (1%) pericytes synthesize and release cyclopentenone prostaglandins. Increased PGD2 and the dehydration product delta12pGJ2 were detected by HPLC and by immune techniques. Delta12pGD2 was not synthesized. Pericytes constitutively express three of the five alternate splice variants of vascular endothelial cell growth factor (VEGF) mRNA, but little to no protein. Exposure to low oxygen increased mRNA levels as well as the synthesis and release of VEGF protein. Addition of either Delta12pGJ2 or 15-deoxyDelta12/14PGJ2 to pericyte under normoxic conditions increased the synthesis and release of VEGF protein in a dose dependent manner. Results suggest that PGD produced by the CNS pericyte is an early signaling molecule in regulation of the angiogenic response to hypoxia. We will further investigate the pericyte VEGF response:
Aim # l: Determine the mechanism of hypoxia induced upregulation of VEGF gene expression. We will also evaluate the role of translation initially and inhibiting factor elF-4E and 4EBprincipal investigator.
Aim#2 : To determine the role of COX-1 in pericyte VEGF response to hypoxia.
Aim#3 : To determine the role of the COX-1 mediated release of pericyte PGD/PGJ in hypoxia. We propose that the CNS microvascular pericyte is a regulatory cell important in the vascular response to hypoxic stress and that cyclopentenone prostagladins are involved in the pericyte stress response.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS047672-01
Application #
6719501
Study Section
Special Emphasis Panel (ZRG1-BDCN-3 (02))
Program Officer
Jacobs, Tom P
Project Start
2004-01-01
Project End
2007-12-31
Budget Start
2004-01-01
Budget End
2004-12-31
Support Year
1
Fiscal Year
2004
Total Cost
$335,035
Indirect Cost
Name
Wayne State University
Department
Neurology
Type
Schools of Medicine
DUNS #
001962224
City
Detroit
State
MI
Country
United States
Zip Code
48202
Katyshev, Vladimir; Dore-Duffy, Paula (2012) Pericyte coculture models to study astrocyte, pericyte, and endothelial cell interactions. Methods Mol Biol 814:467-81
Dore-Duffy, Paula; Mehedi, Afroza; Wang, Xueqian et al. (2011) Immortalized CNS pericytes are quiescent smooth muscle actin-negative and pluripotent. Microvasc Res 82:18-27
Li, Longxuan; Welser, Jennifer V; Dore-Duffy, Paula et al. (2010) In the hypoxic central nervous system, endothelial cell proliferation is followed by astrocyte activation, proliferation, and increased expression of the alpha 6 beta 4 integrin and dystroglycan. Glia 58:1157-67
Milner, Richard; Hung, Stephanie; Erokwu, Bernadette et al. (2008) Increased expression of fibronectin and the alpha 5 beta 1 integrin in angiogenic cerebral blood vessels of mice subject to hypobaric hypoxia. Mol Cell Neurosci 38:43-52
Dore-Duffy, Paula; Katychev, Andre; Wang, Xueqian et al. (2006) CNS microvascular pericytes exhibit multipotential stem cell activity. J Cereb Blood Flow Metab 26:613-24
Dore-Duffy, Paula; Balabanov, Roumen; Beaumont, Thomas et al. (2005) The CNS pericyte response to low oxygen: early synthesis of cyclopentenone prostaglandins of the J-series. Microvasc Res 69:79-88