This proposal aims to examine, for the first time, the correlation between endothelial ultrastructural differentiation and macromolecular permselectivity during normal angiogenesis. In particular, the role of biomolecular differentiation of the endothelial glycocalyx in modulating the access of macromolecules to the junctional clefts and plasmalemmal vesicles, during their concomitant ultrastructural differentiation, will be evaluated. The extraembryonic microcirculation of the chick chorioallantoic membrane (CAM), which requires neither anesthesia nor surgical manipulation for direct microscopic observation, will serve as models of angiogenesis. Evaluations of precapillary, capillary, and postcapillary segments at days 6, 10, 14, and 18 of embryogenesis will be approached, in part, by intravital fluorescence microscopy and the integrated optical intensity technique to assess differential rates of extravasation of a series of graded molecular weight FITC-dextrans. The measured rates will provide an index of differentiation of endothelial permselectivity during normal angiogenesis. Ultrastructural morphometric detection of these same dextrans by colloidal gold immunocytochemistry will serve to correlate differentiation of endothelial plasmalemmal vesicles and junctional clefts with macromolecular permselectivity. Ultrastructural profiles of specific lectins and charged markers within the endothelial glycocalyx during the stages of endothelial cytodifferentiation will complete the analysis of the paracellular and vesicular pathways. Thus, the influence of biomolecular differentiation of the glucocalyx on the regulation of macromolecular access to these pathways will be evaluated. In addition, the effects of histamine on the glycoconjugate microdomains within the glycocalyx, and hence, on macromolecular transport will be tested. Taken as a whole, these data on normal angiogenesis will serve to provide the basis for understanding the derangements that occur during pathologic angiogenesis associated with wound healing and tumorigenesis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL047936-05
Application #
2685388
Study Section
Pathology A Study Section (PTHA)
Project Start
1992-02-01
Project End
2000-03-31
Budget Start
1998-04-01
Budget End
1999-03-31
Support Year
5
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Medicine & Dentistry of NJ
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
605799469
City
Newark
State
NJ
Country
United States
Zip Code
07107
Defouw, L M; Defouw, D O (2001) Protein kinase C activity contributes to endothelial hyperpermeability during early angiogenesis in the chick chorioallantoic membrane. Tissue Cell 33:135-40
DeFouw, L M; DeFouw, D O (2001) Differential phosphodiesterase activity contributes to restrictive endothelial barrier function during angiogenesis. Microvasc Res 62:263-70
DeFouw, L M; DeFouw, D O (2000) Differentiation of endothelial barrier function during normal angiogenesis requires homotypic VE-cadherin adhesion. Tissue Cell 32:238-42
Kissin, M; Kansal, N; Pappas, P J et al. (2000) Vein interposition cuffs decrease the intimal hyperplastic response of polytetrafluoroethylene bypass grafts. J Vasc Surg 31:69-83
DeFouw, L M; DeFouw, D O (2000) Vascular endothelial growth factor fails to acutely modulate endothelial permeability during early angiogenesis in the chick chorioallantoic membrane. Microvasc Res 60:212-21
Defouw, L M; Defouw, D O (2000) Cyclic GMP-mediated macromolecular extravasation from angiogenic microvessels in vivo. Endothelium 7:193-200
Cruz, A; DeFouw, L M; DeFouw, D O (2000) Restrictive endothelial barrier function during normal angiogenesis in vivo: partial dependence on tyrosine dephosphorylation of beta-catenin. Microvasc Res 59:195-203
Cruz, A; DeFouw, D O (1999) Increased expression of VE-cadherin correlates temporally with differentiation of a restrictive endothelial barrier during normal angiogenesis in vivo. Tissue Cell 31:545-9
DeFouw, L M; DeFouw, D O (1999) Modulation of angiogenic endothelial permselectivity by the cAMP pathway. Microvasc Res 57:19-29
Pappas, P J; Gwertzman, G A; DeFouw, D O et al. (1998) Retinoblastoma protein: a molecular regulator of chronic venous insufficiency. J Surg Res 76:149-53

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