Chemically Induced Cell Fusion in vitro of Erythrocytes from Patients with Liver Diseases

1977 ◽  
Vol 5 (4) ◽  
pp. 1144-1146 ◽  
Author(s):  
MICHAEL J. HOPE ◽  
K. RICHARD BRUCKDORFER ◽  
JAMES S. OWEN ◽  
JACK A. LUCY
Keyword(s):  
Cell Fusion ◽  
Liver Diseases ◽  
Chemically Induced

Human 3-dimensional liver organoids: in vitro model systems to study liver diseases

2020 ◽  
Author(s):  
H Gaitantzi ◽  
C Cai ◽  
S Asawa ◽  
K Böttcher ◽  
M Ebert ◽  
...  
Keyword(s):  
Liver Diseases ◽  
In Vitro Model ◽  
Model Systems ◽  
3 Dimensional ◽  
Vitro Model ◽  
Liver Organoids

scholarly journals The Chlamydomonas Mating Type Plus Fertilization Tubule, a Prototypic Cell Fusion Organelle: Isolation, Characterization, and In Vitro Adhesion to Mating Type Minus Gametes

1997 ◽  
Vol 137 (7) ◽  
pp. 1537-1553 ◽  
Author(s):  
Nedra F. Wilson ◽  
Mary J. Foglesong ◽  
William J. Snell
Keyword(s):  
Cell Fusion ◽  
Mating Type ◽  
Plasma Membranes ◽  
Surface Proteins ◽  
Filamentous Actin ◽  
Mating Structure ◽  
Cell Protrusion ◽  
Isolation And Characterization ◽  
In Vitro Adhesion

In the biflagellated alga Chlamydomonas, adhesion and fusion of the plasma membranes of gametes during fertilization occurs via an actin-filled, microvillus-like cell protrusion. Formation of this ∼3-μm-long fusion organelle, the Chlamydomonas fertilization tubule, is induced in mating type plus (mt+) gametes during flagellar adhesion with mating type minus (mt−) gametes. Subsequent adhesion between the tip of the mt+ fertilization tubule and the apex of a mating structure on mt− gametes is followed rapidly by fusion of the plasma membranes and zygote formation. In this report, we describe the isolation and characterization of fertilization tubules from mt+ gametes activated for cell fusion. Fertilization tubules were detached by homogenization of activated mt+ gametes in an EGTA-containing buffer and purified by differential centrifugation followed by fractionation on sucrose and Percoll gradients. As determined by fluorescence microscopy of samples stained with a fluorescent probe for filamentous actin, the method yielded 2–3 × 106 fertilization tubules/μg protein, representing up to a 360-fold enrichment of these organelles. Examination by negative stain electron microscopy demonstrated that the purified fertilization tubules were morphologically indistinguishable from fertilization tubules on intact, activated mt+ gametes, retaining both the extracellular fringe and the internal array of actin filaments. Several proteins, including actin as well as two surface proteins identified by biotinylation studies, copurified with the fertilization tubules. Most importantly, the isolated mt+ fertilization tubules bound to the apical ends of activated mt− gametes between the two flagella, the site of the mt− mating structure; a single fertilization tubule bound per cell, binding was specific for gametes, and fertilization tubules isolated from trypsin-treated, activated mt+ gametes did not bind to activated mt− gametes.

scholarly journals Developmental Neurotoxicity of Environmentally Relevant Pharmaceuticals and Mixtures Thereof in a Zebrafish Embryo Behavioural Test

2021 ◽  
Vol 18 (13) ◽  
pp. 6717
Author(s):  
Alessandro Atzei ◽  
Ingrid Jense ◽  
Edwin P. Zwart ◽  
Jessica Legradi ◽  
Bastiaan J. Venhuis ◽  
...  
Keyword(s):  
Locomotor Activity ◽  
Animal Studies ◽  
Zebrafish Embryo ◽  
Cumulative Effect ◽  
Developmental Neurotoxicity ◽  
Limited Information ◽  
Behavioural Phenotype ◽  
Chemically Induced ◽  
Behavioural Test

Humans are exposed daily to complex mixtures of chemical substances via food intake, inhalation, and dermal contact. Developmental neurotoxicity is an understudied area and entails one of the most complex areas in toxicology. Animal studies for developmental neurotoxicity (DNT) are hardly performed in the context of regular hazard studies, as they are costly and time consuming and provide only limited information as to human relevance. There is a need for a combination of in vitro and in silico tests for the assessment of chemically induced DNT in humans. The zebrafish (Danio rerio) embryo (ZFE) provides a powerful model to study DNT because it shows fast neurodevelopment with a large resemblance to the higher vertebrate, including the human system. One of the suitable readouts for DNT testing in the zebrafish is neurobehaviour (stimulus-provoked locomotion) since this provides integrated information on the functionality and status of the entire nervous system of the embryo. In the current study, environmentally relevant pharmaceuticals and their mixtures were investigated using the zebrafish light-dark transition test. Zebrafish embryos were exposed to three neuroactive compounds of concern, carbamazepine (CBZ), fluoxetine (FLX), and venlafaxine (VNX), as well as their main metabolites, carbamazepine 10,11-epoxide (CBZ 10,11E), norfluoxetine (norFLX), and desvenlafaxine (desVNX). All the studied compounds, except CBZ 10,11E, dose-dependently inhibited zebrafish locomotor activity, providing a distinct behavioural phenotype. Mixture experiments with these pharmaceuticals identified that dose addition was confirmed for all the studied binary mixtures (CBZ-FLX, CBZ-VNX, and VNX-FLX), thereby supporting the zebrafish embryo as a model for studying the cumulative effect of chemical mixtures in DNT. This study shows that pharmaceuticals and a mixture thereof affect locomotor activity in zebrafish. The test is directly applicable in environmental risk assessment; however, further studies are required to assess the relevance of these findings for developmental neurotoxicity in humans.

scholarly journals Disulfide Bond Formation at the C Termini of Vaccinia Virus A26 and A27 Proteins Does Not Require Viral Redox Enzymes and Suppresses Glycosaminoglycan-Mediated Cell Fusion

2009 ◽  
Vol 83 (13) ◽  
pp. 6464-6476 ◽  
Author(s):  
Yao-Cheng Ching ◽  
Che-Sheng Chung ◽  
Cheng-Yen Huang ◽  
Yu Hsia ◽  
Yin-Liang Tang ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Cell Fusion ◽  
Disulfide Bond ◽  
Protein Complex ◽  
Disulfide Bonds ◽  
Bond Formation ◽  
In Vitro Mutagenesis ◽  
Disulfide Bond Formation ◽  
Infected Cells

ABSTRACT Vaccinia virus A26 protein is an envelope protein of the intracellular mature virus (IMV) of vaccinia virus. A mutant A26 protein with a truncation of the 74 C-terminal amino acids was expressed in infected cells but failed to be incorporated into IMV (W. L. Chiu, C. L. Lin, M. H. Yang, D. L. Tzou, and W. Chang, J. Virol 81:2149-2157, 2007). Here, we demonstrate that A27 protein formed a protein complex with the full-length form but not with the truncated form of A26 protein in infected cells as well as in IMV. The formation of the A26-A27 protein complex occurred prior to virion assembly and did not require another A27-binding protein, A17 protein, in the infected cells. A26 protein contains six cysteine residues, and in vitro mutagenesis showed that Cys441 and Cys442 mediated intermolecular disulfide bonds with Cys71 and Cys72 of viral A27 protein, whereas Cys43 and Cys342 mediated intramolecular disulfide bonds. A26 and A27 proteins formed disulfide-linked complexes in transfected 293T cells, showing that the intermolecular disulfide bond formation did not depend on viral redox pathways. Finally, using cell fusion from within and fusion from without, we demonstrate that cell surface glycosaminoglycan is important for virus-cell fusion and that A26 protein, by forming complexes with A27 protein, partially suppresses fusion.

scholarly journals Molecular characterization of the grape seeds extract’s effect against chemically induced liver cancer: In vivo and in vitro analyses

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Alaaeldin Ahmed Hamza ◽  
Gehan Hussein Heeba ◽  
Hanan Mohamed Elwy ◽  
Chandraprabha Murali ◽  
Raafat El-Awady ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Molecular Characterization ◽  
Grape Seeds ◽  
Chemically Induced

scholarly journals Investigating FlowSight® imaging flow cytometry as a platform to assess chemically induced micronuclei using human lymphoblastoid cells in vitro

Mutagenesis ◽  
2018 ◽  
Vol 33 (4) ◽  
pp. 283-289 ◽  
Author(s):  
Jatin R Verma ◽  
Danielle S G Harte ◽  
Ume-Kulsoom Shah ◽  
Huw Summers ◽  
Catherine A Thornton ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Lymphoblastoid Cells ◽  
Imaging Flow Cytometry ◽  
Chemically Induced

Studies on Chemically Induced Cell Fusion

1972 ◽  
Vol 10 (3) ◽  
pp. 769-787
Author(s):  
Q. F. AHKONG ◽  
F. C. CRAMP ◽  
D. FISHER ◽  
J. I. HOWELL ◽  
J. A. LUCY
Keyword(s):  
Aqueous Solution ◽  
Cell Fusion ◽  
Plasma Membranes ◽  
Mouse Fibroblast ◽  
Nuclear Membranes ◽  
Multinucleated Cells ◽  
Large Numbers ◽  
Chemically Induced ◽  
Complete Disintegration ◽  
Unstable Plasma

Hen erythrocytes that were fixed after treatment with lysolecithin in aqueous solution for 30 s at 37 °C showed evidence of bridge formation between adjacent lysed cells. Generally, the homokaryons that were produced using lysolecithin in this way contained large numbers of nuclei. These giant syncytia had damaged nuclear membranes and unstable plasma membranes; complete disintegration of the syncytia occurred within 1 min of adding lysolecithin to the erythrocytes. In order to localize the action of lysolecithin, the fusing agent was incorporated into microdroplets of lipid. Cell fusion following the addition of lysolecithin in an aqueous glyceridelecithin emulsion was slower than with lysolecithin in aqueous solution, taking 10-30 min, and it was accompanied by considerably less damage to the plasma and nuclear membranes. The fused erythrocytes, which usually contained only two or three nuclei, lysed slowly during the 45 min following fusion, and lysis could be arrested by cooling the fused cells. The plasma membranes of lysed, multinucleated cells remained intact at 37°C for at least 90 h. Mouse fibroblast-hen erythrocyte heterokaryons formed with the aid of the emulsion were more stable than those produced with lysolecithin in solution, but the hybrid cells nevertheless had damaged subcellular organelles. Viable clones of hybrid mouse-hamster fibroblast cells were obtained using the emulsion although, possibly owing to reduced viability of the lysolecithin-treated cells, only at twice the frequency of spontaneously produced hybrids.

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