Endothelial Glycocalyx-Mediated Nitric Oxide Production in Response to Selective AFM Pulling

被引:83
作者
Bartosch, Anne Marie W. [1 ]
Mathews, Rick [1 ,2 ]
Tarbell, John M. [1 ]
机构
[1] CUNY, Dept Biomed Engn, New York, NY 10021 USA
[2] William E Macaulay Honors Coll, New York, NY USA
基金
美国国家卫生研究院;
关键词
FLUID SHEAR-STRESS; HEPARAN-SULFATE; RHAMNAN SULFATE; ARTERY; MECHANOTRANSDUCTION; GLYCOSAMINOGLYCANS; TRANSMISSION; COMBINATION; GLYPICAN-1; RECEPTOR;
D O I
10.1016/j.bpj.2017.05.033
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Nitric oxide (NO) is a regulatory molecule in the vascular system and its inhibition due to endothelial injury contributes to cardiovascular disease. The glycocalyx is a thin layer of glycolipids, glycoproteins, and proteoglycans on the surface of mammalian epithelial cells. Extracellular forces are transmitted through the glycocalyx to initiate intracellular signaling pathways. In endothelial cells (ECs), previous studies have shown the glycocalyx to be a significant mediator of NO production; degradation of the endothelial glycocalyx layer (EGL) drastically reduces EC production of NO in response to fluid shear stress. However, the specific EGL components involved in this process are not well established. Recent work using short-hairpin RNA approaches in vitro suggest that the proteoglycan glypican-1, not syndecan-1, is the dominant core protein mediating shear induced NO production. We utilized atomic force microscopy (AFM) to apply force selectively to components of the EGL of confluent rat fat pad ECs (RFPECs), including proteoglycans and glycosaminoglycans, to observe how each component individually contributes to force-induced production of NO. 4,5-diaminofluorescein diacetate, a cell-permeable fluorescent molecule, was used to detect changes in intracellular NO production. Antibody-coated AFM probes exhibited strong surface binding to RFPEC monolayers, with 100-300 pN mean adhesion forces. AFM pulling on glypican-1 and heparan sulfate for 10 min caused significantly increased NO production, whereas pulling on syndecan-1, CD44, hyaluronic acid, and with control probes did not. We conclude that AFM pulling can be used to activate EGL-mediated NO production and that the heparan sulfate proteoglycan glypican-1 is a primary mechanosensor for shear-induced NO production.
引用
收藏
页码:101 / 108
页数:8
相关论文
共 42 条
[1]   CALCULATION OF THERMAL NOISE IN ATOMIC-FORCE MICROSCOPY [J].
BUTT, HJ ;
JASCHKE, M .
NANOTECHNOLOGY, 1995, 6 (01) :1-7
[2]   Platelet endothelial cell adhesion molecule 1 (PECAM-1) and its interactions with glycosaminoglycans: 2. Biochemical analyses [J].
Coombe, Deirdre R. ;
Stevenson, Sandra M. ;
Kinnear, Beverley F. ;
Gandhi, Neha S. ;
Mancera, Ricardo L. ;
Osmond, Ronald I. W. ;
Kettt, Warren C. .
BIOCHEMISTRY, 2008, 47 (17) :4863-4875
[3]   Heparan Sulfates Mediate the Interaction between Platelet Endothelial Cell Adhesion Molecule-1 (PECAM-1) and the Gαq/11 Subunits of Heterotrimeric G Proteins [J].
dela Paz, Nathaniel G. ;
Melchior, Benoit ;
Shayo, Francisca Y. ;
Frangos, John A. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2014, 289 (11) :7413-7424
[4]   PECAM-1 interacts with nitric oxide synthase in human endothelial cells - Implication for flow-induced nitric oxide synthase activation [J].
Dusserre, N ;
L'Heureux, N ;
Bell, KS ;
Stevens, HY ;
Yeh, J ;
Otte, LA ;
Loufrani, L ;
Frangos, JA .
ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2004, 24 (10) :1796-1802
[5]   Functionalization of probe tips and supports for single-molecule recognition force Microscopy [J].
Ebner, Andreas ;
Wildling, Linda ;
Zhu, Rong ;
Rankl, Christian ;
Haselgruebler, Thomas ;
Hinterdorfer, Peter ;
Gruber, Hermann J. .
STM AND AFM STUDIES ON (BIO)MOLECULAR SYSTEMS: UNRAVELLING THE NANOWORLD, 2008, 285 :29-76
[6]   Shear-induced endothelial NOS activation and remodeling via heparan sulfate, glypican-1, and syndecan-1 [J].
Ebong, Eno E. ;
Lopez-Quintero, Sandra V. ;
Rizzo, Victor ;
Spray, David C. ;
Tarbell, John M. .
INTEGRATIVE BIOLOGY, 2014, 6 (03) :338-347
[7]   Imaging the Endothelial Glycocalyx In Vitro by Rapid Freezing/Freeze Substitution Transmission Electron Microscopy [J].
Ebong, Eno E. ;
Macaluso, Frank P. ;
Spray, David C. ;
Tarbell, John M. .
ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 2011, 31 (08) :1908-1915
[8]   Heparan sulfate proteoglycan is a mechanosensor on endothelial cells [J].
Florian, JA ;
Kosky, JR ;
Ainslie, K ;
Pang, ZY ;
Dull, RO ;
Tarbell, JM .
CIRCULATION RESEARCH, 2003, 93 (10) :E136-E142
[9]   Mechano-sensing and transduction by endothelial surface glycocalyx: composition, structure, and function [J].
Fu, Bingmei M. ;
Tarbell, John M. .
WILEY INTERDISCIPLINARY REVIEWS-SYSTEMS BIOLOGY AND MEDICINE, 2013, 5 (03) :381-390
[10]   NITRIC OXIDE-GENERATING VASODILATORS AND 8-BROMO-CYCLIC GUANOSINE-MONOPHOSPHATE INHIBIT MITOGENESIS AND PROLIFERATION OF CULTURED RAT VASCULAR SMOOTH-MUSCLE CELLS [J].
GARG, UC ;
HASSID, A .
JOURNAL OF CLINICAL INVESTIGATION, 1989, 83 (05) :1774-1777