A New Model for Quantitative In Vivo Microscopic Analysis of Thrombus Formation and Vascular Recanalisation: The Ear of the Hairless (hr/hr) Mouse

SummaryThe alteration of rheological blood properties as well as deterioration of vascular perfusion conditions and cell-cell interactions are major determinants of thrombus formation. Herein, we present an experimental model which allows for quantitative in vivo microscopic analysis of these determinants during both thrombus formation and vascular recanalisation. The model does not require surgical preparation procedures, and enables for repeated analysis of identical microvessels over time periods of days or months, respectively. After i.v. administration of FITC-dextran thrombus formation was induced photochemically by light exposure to individual arterioles and venules of the ear of ten anaesthetised hairless mice. In venules, epiillumination induced rapid thrombus formation with first platelet deposition after 0.59 ± 0.04 min and complete vessel occlusion within 7.48 ±1.31 min. After a 24-h time period, 75% of the thrombosed venules were found recanalised. Marked leukocyte-endothelial cell interaction in those venules indicated persistent endothelial cell activation and/or injury, even after an observation period of 7 days. In arterioles, epi-illumination provoked vasomotion, while thrombus formation was significantly (p <0.05) delayed with first platelet deposition after 2.32 ± 0.22 min and complete vessel occlusion within 20.07 ±3.84 min. Strikingly, only one of the investigated arterioles was found recanalised after 24 h, which, however, did not show leukocyte-endothelial cell interaction. Heparin (300 U/kg, i.v.) effectively counteracted the process of thrombus formation in this model, including both first platelet deposition and vessel occlusion. We conclude that the model of the ear of the hairless mouse allows for distinct in vivo analysis of arteriolar and venular thrombus formation/ recanalisation, and, thus, represents an interesting tool for the study of novel antithrombotic and thrombolytic strategies, respectively.

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Lack of In Vivo Function of CD31 in Vascular Thrombosis

Thrombosis and Haemostasis ◽

10.1055/s-0037-1612919 ◽

2001 ◽

Vol 85 (01) ◽

pp. 160-164 ◽

Cited By ~ 34

Author(s):

R. Schmits ◽

D. Kunz ◽

M. D. Menger ◽

B. Vollmar

Keyword(s):

Endothelial Cell ◽

Blood Cell ◽

Cell Injury ◽

Thrombus Formation ◽

Vessel Occlusion ◽

Endothelial Cell Injury ◽

Vascular Thrombosis ◽

Platelet Deposition ◽

Deficient Mice

SummaryA murine model of endothelial cell injury-based vascular thrombosis was used to test the role of platelet-endothelial cell adhesion molecule-1 (PECAM-1, CD31) in blood cell aggregate formation and vessel occlusion in vivo. Photochemically-induced thrombus formation was analyzed in detail using intravital fluorescence microscopy of individual microvessels in cremaster muscle preparations of CD31-deficient and wildtype mice. In venules, epi-illumination induced rapid thrombus formation with first platelet deposition after 0.56 ± 0.11min and complete vessel occlusion within 5.05 ± 0.45 min. In arterioles, thrombus formation was markedly delayed with first platelet deposition after 3.03 ± 0.47 min and complete vessel occlusion within 10.04 ± 1.26 min. Kinetics of thrombus formation in both venules (first platelet deposition: 0.52 ± 0.1 min; vessel occlusion: 5.03 ± 0.52 min) and arterioles (first platelet deposition: 3.06 ± 0.68 min; vessel occlusion: 10.02 ± 1.38 min) of CD31-deficient mice was found almost identical compared with that in wildtype animals. Tail bleeding time was 233 ± 24 s in wildtype and 243 ± 32 s in CD31-deficient mice. Moreover, CD31-deficient and wildtype mice revealed comparable interaction of leukocytes to endothelium. This study shows for the first time in vivo that CD31 is not critically involved in blood cell thrombus formation upon endothelial cell injury.

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In Vivo Assessment of Synovial Microcirculation and Leukocyte–Endothelial Cell Interaction in Mouse Antigen-Induced Arthritis

Microcirculation ◽

10.1038/sj.mn.7300074 ◽

1999 ◽

Vol 6 (4) ◽

pp. 281-290 ◽

Cited By ~ 1

Author(s):

A N D R E A S VEIHELMANN ◽

ANTHONY G U S T A V E HARRIS ◽

F R I T Z KROMBACH ◽

E L K E SCHÜTZE ◽

HANS JÜRGEN REFIOR ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Interaction ◽

Endothelial Cell Interaction ◽

In Vivo Assessment

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In vivo that intercellular adhesion molecule-1 does not mediate endotoxin-induced hepatic leukocyte-endothelial cell interaction

Journal of Hepatology ◽

10.1016/0168-8278(95)80070-0 ◽

1995 ◽

Vol 23 (5) ◽

pp. 613-616 ◽

Cited By ~ 15

Author(s):

Brigitte Vollmar ◽

Achmed Senkel ◽

Michael D. Menger

Keyword(s):

Endothelial Cell ◽

Adhesion Molecule ◽

Cell Interaction ◽

Intercellular Adhesion ◽

Intercellular Adhesion Molecule ◽

Intercellular Adhesion Molecule 1 ◽

Endothelial Cell Interaction

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Genetic control of leucocyte—endothelial cell interaction in collagen-induced arthritis

Annals of the Rheumatic Diseases ◽

10.1136/ard.2009.100636 ◽

2010 ◽

Vol 69 (3) ◽

pp. 606-610 ◽

Cited By ~ 1

Author(s):

Hoang Tu-Rapp ◽

Liying Pu ◽

Andreia Marques ◽

Christoph Kulisch ◽

Xinhua Yu ◽

...

Keyword(s):

Endothelial Cell ◽

Tissue Injury ◽

Genome Scan ◽

Cell Interaction ◽

Collagen Induced Arthritis ◽

Endothelial Lining ◽

Phenotypic Data ◽

Endothelial Cell Interaction ◽

A Genome

ObjectiveDespite considerable work on defining disease pathways, several aspects of collagen-induced arthritis (CIA) remain poorly defined, in particular those contributing to the initiation phase of the disease. It is thought that in CIA the activation of circulating leucocytes, their interaction with the endothelial lining followed by subsequent transendothelial migration and infiltration into tissue represents the first and determining step in a complex sequence of processes mediating tissue injury. In this study we attempted to define the genetic basis of this stage of disease using genetic linkage studies, in-vivo imaging and expression profiling.MethodsA genome scan with 132 informative markers was performed on 155 (DBA/1J×FVB/N) F2 mice. Linkage analysis was performed by combining genotyping data from the genome scan and the phenotypic data of leucocyte adherence, leucocyte rolling fraction, functional capillary density, centre line red blood cell velocity and capillary width as well as the expression level of the selected genes Cd44, Il13rα1, Ccr3, Defb3, Sele, Sell, Selp, Xcl1, Il1β, Tnfα and Ifnγ as traits.ResultsMultiple classic quantitative trail loci (QTL) controlling leucocyte–endothelial cell interactions were identified on chromosomes 8 and 17 as well as expression QTL controlling the expression of several differentially expressed adhesion molecules and cytokines on chromosomes 1, 2, 5, 6, 7, 8, 12, 15, 16 and 17.ConclusionThe study describes for the first time QTL controlling the CIA initiating leucocyte–endothelial cell interaction.

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In Vivo Assessment of Synovial Microcirculation and Leukocyte-Endothelial Cell Interaction in Mouse Antigen-Induced Arthritis

Microcirculation ◽

10.1080/713773963 ◽

1999 ◽

Vol 6 (4) ◽

pp. 281-290

Author(s):

Andreas Veihelmann ◽

Anthony Gustave Harris ◽

Fritz Krombach ◽

Elke Schütze ◽

Hans Jürgen Refior ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Interaction ◽

Endothelial Cell Interaction ◽

In Vivo Assessment

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In vivo leucocyte-endothelial cell interaction induced by extracorporeal circulation: reduction by a coated tube system

Critical Care ◽

10.1186/cc315 ◽

1999 ◽

Vol 3 (S2) ◽

Author(s):

M Kamler ◽

T Chatterjee ◽

H Jakob ◽

A Stemberger ◽

MM Gebhard ◽

...

Keyword(s):

Endothelial Cell ◽

Extracorporeal Circulation ◽

Cell Interaction ◽

Tube System ◽

Endothelial Cell Interaction

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Endothelial iNOS versus platelet iNOS: Responsibility for the platelet/leukocyte endothelial cell interaction in murine antigen induced arthritis in vivo

Inflammation Research ◽

10.1007/s00011-007-6171-x ◽

2007 ◽

Vol 56 (6) ◽

pp. 262-268 ◽

Cited By ~ 6

Author(s):

M. Schmitt-Sody ◽

O. Gottschalk ◽

P. Metz ◽

S. Zysk ◽

J. Hausdorf ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Interaction ◽

Endothelial Cell Interaction

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Platelet-endothelial cell interaction in pulmonary microcirculation: the role of PARS

Thrombosis and Haemostasis ◽

10.1160/th03-11-0685 ◽

2004 ◽

Vol 91 (04) ◽

pp. 761-770 ◽

Cited By ~ 19

Author(s):

Rainer Kiefmann ◽

Kai Heckel ◽

Sonja Schenkat ◽

Martina Dörger ◽

Józefa Węsierska-Gądek ◽

...

Keyword(s):

Endothelial Cell ◽

Cell Interaction ◽

Microvascular Endothelial Cells ◽

Microvascular Endothelium ◽

Endotoxin Infusion ◽

Endothelial Cell Interaction ◽

Pulmonary Microcirculation ◽

Kinetics Of

SummaryAccumulation of platelets might contribute to acute lung injury during systemic inflammation. The aim of the study was to elucidate the role of the poly (ADP-ribose) synthetase, a nucleotide-polymerizising enzyme, in mediation of platelet-endothelial cell interaction through regulation of adhesion molecules within the pulmonary microcirculation during endotoxemia. We used in vivo fluorescence microscopy to quantify kinetics of fluorescently labeled erythrocytes and platelets in rabbit pulmonary arterioles and venules. Six hours after onset of endotoxin infusion we observed a massive interaction of platelets with the microvascular endothelial cells, whereas under control conditions, no platelet sequestration was measured. An up-regulation of P- and E-selectin was detected in lung tissue following endotoxin infusion by immunohistochemistry and Western blot analysis. Blockade of endothelial P-selectin with fucoidin resulted in a reduction of the endotoxin-induced platelet-endothelial cell interaction. Inhibition of poly (ADP-ribose) synthetase by 3-aminobenzamide inhibited the endotoxin-induced expression of endothelial P- and E-selectin and the subsequent recruitment of platelets. In summary, we provide first in vivo evidence that platelets accumulate in pulmonary microcirculation following endotoxemia. Poly (ADP-ribose) synthetase seems to mediate this platelet-endothelial cell interaction via P- and E-selectin expressed on the surface of microvascular endothelium.

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Heparin - PF4 Pulsing as a Measure of Platelet - Endothelial Cell Reaction in Vivo

10.1055/s-0039-1684365 ◽

1979 ◽

Author(s):

C.W. Pumphrey ◽

D.S. Pepper ◽

J. Dawes

Keyword(s):

Endothelial Cell ◽

Vascular Endothelium ◽

Cell Interaction ◽

Cell Reaction ◽

Heparin Dose ◽

Normal Value ◽

Endothelial Cell Interaction

iv, Heparin (1000 units) causes an increase in plasma, PF4 of as much as 30-fold, when treasured by radioiiumunoassay. The PF4 reaches a peak within 5 minutes after injection and Is subsequently cleared to a normal value within 40-80 minutes. The Peak elevation is proportional to the initial heparin dose. Subsequent iv, Heparin doses, within the next few hours, fail to elevate PF4. Control experiments strongly indicate that the source of the PF4 is the vascular endothelium rather than the platelets. Further experiments have shown that this PF4. reservoir half fills within 46 hours. We suggest that iv. heparin pulses can be used for studying, indirectly, platelet - endothelial cell interaction in vivo.

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