Preparation of tetramethylrhodaminyl-phalloidin and uptake of the toxin into short-term cultured hepatocytes by endocytosis

Faulstich, Heinz; Trischmann, Heinrich; Mayer, Doris

doi:10.1016/0014-4827(83)90443-3

Cited by 115 publications

(33 citation statements)

References 9 publications

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“…Afterwards each coverslip was incubated with 500 µl (1 µg/ml) phalloidin covalently labelled with tetramethyl-rhodamine isothiocyanate (TRITC) for 1 hour at 37°C (Faulstich et al, 1983;Franke et al, 1984). Series of experiments with changing concentrations of TRITC-phalloidin (0.1-10 µM) were conducted to show that the conditions chosen allow saturation of binding.…”

Section: Quantification Of F-actin and Ve-cadherinmentioning

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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Section: Quantification Of F-actin and Ve-cadherinmentioning

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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“…For determination of the relative F-actin contents, PC12 cells were fixed at room temperature with 3% formaldehyde in PBS for 15 min and then permeabilized with 0.1% (v/v) Triton X-100 in PBS for 5 min. Afterward, each coverslip was incubated with 500 l (1 g/ml) of phalloidin covalently labeled with tetramethylrhodamine isothiocyanate (TRITC) for 1 hr at 37°C (Faulstich et al, 1983). Series of experiments with changing concentrations of TRITC-phalloidin (0.1-10 M) were conducted to show that the conditions chosen allowed saturation of binding.…”

Section: Methodsmentioning

confidence: 99%

Ca²⁺Dependency of N-Cadherin Function Probed by Laser Tweezer and Atomic Force Microscopy

et al. 2003

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“…The mean value of each parameter was determined over a 10-20 min period for each synapse before averaging. Faulstich & Wieland, 1979;Faulstich, Trischmann & Mayer, 1983) after fixation and permeabilization of the cells. Both isolated spinal neurons and neurons that had innervated co-cultured myocytes were examined.…”

Section: Effects Of Cytochalasins On Actin Filamentsmentioning

confidence: 99%

Effects of cytochalasin treatment on short‐term synaptic plasticity at developing neuromuscular junctions in frogs.

Wang

Zheng

Poo

1996

The Journal of Physiology

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1. The role of actin microfilaments in synaptic transmission was tested by monitoring spontaneous and evoked transmitter release from developing neuromuscular synapses in Xenopus nerve-muscle cultures, using whole-cell recording of synaptic currents in the absence and presence of microfilament-disrupting agents cytochalasins B and D. 2. Treatment with cytochalasins resulted in disruption of microfilament networks in the growth cone and the presynaptic nerve terminal of spinal neurons in Xenopus nerve-muscle cultures, as revealed by rhodamine-phalloidin staining. 3. The same cytochalasin treatment did not significantly affect the spontaneous or evoked synaptic currents during low-frequency stimulation at 005 Hz in these Xenopus cultures. Synaptic depression induced by high-frequency (5 Hz) stimulation, however, was reduced by this treatment. Paired-pulse facilitation for short interpulse intervals was also increased by the treatment. 4. These results indicate that disruption of microfilaments alters short-term changes in transmitter release induced by repetitive activity, without affecting normal synaptic transmission at low frequency. 5. Our results support the notion that actin microfilaments impose a barrier for mobilization of synaptic vesicles from the reserve pool, but do not affect the exocytosis of immediately available synaptic vesicles at the active zone.

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Preparation of tetramethylrhodaminyl-phalloidin and uptake of the toxin into short-term cultured hepatocytes by endocytosis

Cited by 115 publications

References 9 publications

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

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

Ca²⁺Dependency of N-Cadherin Function Probed by Laser Tweezer and Atomic Force Microscopy

Effects of cytochalasin treatment on short‐term synaptic plasticity at developing neuromuscular junctions in frogs.

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Preparation of tetramethylrhodaminyl-phalloidin and uptake of the toxin into short-term cultured hepatocytes by endocytosis

Cited by 115 publications

References 9 publications

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

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

Ca2+Dependency of N-Cadherin Function Probed by Laser Tweezer and Atomic Force Microscopy

Effects of cytochalasin treatment on short‐term synaptic plasticity at developing neuromuscular junctions in frogs.

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Ca²⁺Dependency of N-Cadherin Function Probed by Laser Tweezer and Atomic Force Microscopy