Improved in vitro rheological system for studying the effect of fluid shear stress on cultured cells

Schnittler, Hans-J.; Franke, R.P.; Akbay, U.; Mrowietz, C.; Drenckhahn, D.

doi:10.1152/ajpcell.1993.265.1.c289

Cited by 181 publications

(94 citation statements)

References 17 publications

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“…Human embryonic kidney cells (HEK 293, a human fibroblast cell line) were grown on glass coverslips coated with 0.5% gelatine (16) and were transfected with either ARF6-GFP or msec7-1-GFP expression vectors. Alternatively, cells were cotransfected with a combination of either ARF6-GFP͞msec7-1 or ARF6͞ msec7-1-GFP.…”

Section: Methodsmentioning

confidence: 99%

A presynaptic role for the ADP ribosylation factor (ARF)-specific GDP/GTP exchange factor msec7-1

Ashery

Koch

Scheuss

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

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ADP ribosylation factors (ARFs) represent a family of small monomeric G proteins that switch from an inactive, GDP-bound state to an active, GTP-bound state. One member of this family, ARF6, translocates on activation from intracellular compartments to the plasma membrane and has been implicated in regulated exocytosis in neuroendocrine cells. Because GDP release in vivo is rather slow, ARF activation is facilitated by specific guanine nucleotide exchange factors like cytohesin-1 or ARNO. Here we show that msec7-1, a rat homologue of cytohesin-1, translocates ARF6 to the plasma membrane in living cells. Overexpression of msec7-1 leads to an increase in basal synaptic transmission at the Xenopus neuromuscular junction. msec7-1-containing synapses have a 5-fold higher frequency of spontaneous synaptic currents than control synapses. On stimulation, the amplitudes of the resulting evoked postsynaptic currents of msec7-1-overexpressing neurons are increased as well. However, further stimulation leads to a decline in amplitudes approaching the values of control synapses. This transient effect on amplitude is strongly reduced on overexpression of msec7-1E157K, a mutant incapable of translocating ARFs. Our results provide evidence that small G proteins of the ARF family and activating factors like msec7-1 play an important role in synaptic transmission, most likely by making more vesicles available for fusion at the plasma membrane.

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Section: Methodsmentioning

confidence: 99%

A presynaptic role for the ADP ribosylation factor (ARF)-specific GDP/GTP exchange factor msec7-1

Ashery

Koch

Scheuss

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

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“…Cells were grown in Dulbecco's modified Eagle's medium (DMEM; Life Technologies, Eggenstein, Germany) supplemented with 50 U/ml penicillin-G/streptomycin and 10% fetal calf serum (Biochrom) in a humidified atmosphere (95% O2/5% CO2) at 37°C. For experiments cells were grown on coverslips coated with gelatine crosslinked with glutaraldehyde (Schnittler et al, 1993). The culture was split once a week and used for experiments between passages 5 and 20.…”

Section: Cell Culturementioning

confidence: 99%

Cadherin function probed by laser tweezer and single molecule fluorescence in vascular endothelial cells

Baumgärtner

Schütz

Wiegand

et al. 2003

Journal of Cell Science

104

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In endothelial monolayers agonist-induced influx of Ca2+ and activities of the actin cytoskeleton have been shown to be crucially involved in regulation of barrier properties. By laser tweezer application we demonstrated that the strength of adhesion of VE-cadherin-coated microspheres to the surface of cultured endothelial monolayers is significantly reduced by treatment with two well-established permeability-increasing compounds,cytochalasin D and the Ca2+-ionophore A23187, which shows that both compounds directly affect cadherin-mediated adhesion. Cytochalasin D and A23187 caused considerable decay of F-actin (30-60%). Stabilisation of F-actin by jasplakinolide completely blocked drug-induced weakening of bead adhesion showing that attenuation of cadherin-cadherin trans-interaction induced by cytochalasin D and A23187 depends largely on downregulation of F-actin. Single molecule fluorescence microscopy demonstrated that drug-induced weakening of adhesion is accompanied by an increase in lateral mobility of cadherins as well as by dispersal of cadherin-enriched plasmalemmal microdomains. However,the lifetime (≈700 milliseconds, koff≈1.4 second–1) and apparent on-rate of cadherin trans-interaction(relative frequency of binding) remained unchanged in response to cytochalasin D and A23187 indicating that cadherin-mediated adhesion is not modulated by inside-out changes of the affinity but, rather, appears to be controlled by actin-dependent tethering and compartmentalization of cadherins.

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“…In a recent study, an optically transparent CPV system, with multiple rheological units and medium-exchange ports, was designed to investigate the effects of laminar or turbulent fluid shear stress on cultured vascular ECs [63]. Development of laminar and turbulent flow patterns was observed by dissolution of a dye on the plate of the CPV.…”

Section: Rotational Flow Observation and Inertial Effects In Cpvmentioning

confidence: 99%

Biological effects of dynamic shear stress in cardiovascular pathologies and devices

Girdhar

Bluestein

2008

Expert Review of Medical Devices

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Altered and highly dynamic shear stress conditions have been implicated in endothelial dysfunction leading to cardiovascular disease, and in thromboembolic complications in prosthetic cardiovascular devices. In addition to vascular damage, the pathological flow patterns characterizing cardiovascular pathologies and blood flow in prosthetic devices induce shear activation and damage to blood constituents. Investigation of the specific and accentuated effects of such flow-induced perturbations on individual cell-types in vitro is critical for the optimization of device design, whereby specific design modifications can be made to minimize such perturbations. Such effects are also critical in understanding the development of cardiovascular disease. This review addresses limitations to replicate such dynamic flow conditions in vitro and also introduces the idea of modified in vitro devices, one of which is developed in the authors' laboratory, with dynamic capabilities to investigate the aforementioned effects in greater detail.Keywords cardiovascular disease; cone-plate viscometers; dynamic shear stress; platelet activation; prosthetic heart valves; turbulence Implantable blood recirculation devices, artificial hearts and heart valves, have long been used as life-saving alternatives for people with severe cardiovascular diseases. However, such devices require complex anticoagulation therapy and, as a consequence, are linked to postimplant complications, such as hemorrhage. Despite the anticoagulation therapy, the risk for cardioembolic stroke is not eliminated. Although biocompatibility of these recirculation devices is of utmost importance, optimizing the geometric design of these devices is also critical to avoid flow-induced trauma to blood. Over the years, a definite link between fluid shearstress-induced damage and activation of blood constituents such as platelets and red blood cells, has been established. Irregular flow patterns arising around complex geometries (e.g., the hinge regions of mechanical heart valves [MHV]) have been recently characterized by numerical calculations and noninvasive flow measurements, and are implicated in causing flow-induced thromboembolism (TE). For instance, the Medtronic Parallel™ valve exhibited regions of elevated turbulent stress around the hinges due to its specific geometry, which, in †Author for correspondence: Department of Biomedical Engineering, State University of New York at Stony Brook, Stony Brook, NY 11794-8181, USA, Tel.: +1 631 444 1259, Fax: +1 631 444 6646, danny.bluestein@sunysb.edu. Financial & competing interests disclosure:The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties. No writing assistance was utilized in the production of this manu...

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Improved in vitro rheological system for studying the effect of fluid shear stress on cultured cells

Cited by 181 publications

References 17 publications

A presynaptic role for the ADP ribosylation factor (ARF)-specific GDP/GTP exchange factor msec7-1

A presynaptic role for the ADP ribosylation factor (ARF)-specific GDP/GTP exchange factor msec7-1

Cadherin function probed by laser tweezer and single molecule fluorescence in vascular endothelial cells

Biological effects of dynamic shear stress in cardiovascular pathologies and devices

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