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A.M.A. Archives of Internal Medicine Jul 1957
Topics: Colchicine; Demecolcine
PubMed: 13434511
DOI: 10.1001/archinte.1957.00260070099011 -
British Medical Journal Apr 1965
Topics: Colchicine; Demecolcine; Drug Therapy; Leukemia; Neoplasms
PubMed: 14257420
DOI: 10.1136/bmj.1.5439.926-c -
Methods in Molecular Biology (Clifton,... 2016Small molecule drugs that target microtubules (MTs), many of them natural products, have long been important tools in the MT field. Indeed, tubulin (Tb) was discovered,... (Review)
Review
Small molecule drugs that target microtubules (MTs), many of them natural products, have long been important tools in the MT field. Indeed, tubulin (Tb) was discovered, in part, as the protein binding partner of colchicine. Several anti-MT drug classes also have important medical uses, notably colchicine, which is used to treat gout, familial Mediterranean fever (FMF), and pericarditis, and the vinca alkaloids and taxanes, which are used to treat cancer. Anti-MT drugs have in common that they bind specifically to Tb in the dimer, MT or some other form. However, their effects on polymerization dynamics and on the human body differ markedly. Here we briefly review the most-studied molecules, and comment on their uses in basic research and medicine. Our focus is on practical applications of different anti-MT drugs in the laboratory, and key points that users should be aware of when designing experiments. We also touch on interesting unsolved problems, particularly in the area of medical applications. In our opinion, the mechanism by which any MT drug cures or treats any disease is still unsolved, despite decades of research. Solving this problem for particular drug-disease combinations might open new uses for old drugs, or provide insights into novel routes for treatment.
Topics: Animals; Antineoplastic Agents, Phytogenic; Colchicine; Demecolcine; Drug Discovery; Furans; Humans; Ketones; Microtubules; Protein Multimerization; Stilbenes; Sulfonamides; Taxoids; Tubulin Modulators; Vinca Alkaloids
PubMed: 27193863
DOI: 10.1007/978-1-4939-3542-0_25 -
Cellular Reprogramming Oct 2011Demecolcine-assisted/induced enucleation has been used in nuclear transfer cloning procedures for many species, yet its mechanism of action remains unclear. Primarily...
Demecolcine-assisted/induced enucleation has been used in nuclear transfer cloning procedures for many species, yet its mechanism of action remains unclear. Primarily because oocytoplasm protrusion induced by demecolcine is inhibited by the presence of cytochalasin, its use has had limited application. In this experiment, we investigated the microtubule and microfilament alterations in bovine oocytes after demecolcine and/or cytochalasin B (CB) treatments by immunocytochemical staining. We also examined mechanical enucleation of demecolcine-treated oocytes in cytochalasin-free medium. The results showed that demecolcine-treatment disrupts the balance between microtubule/microfilament interactions primarily by deleting microtubules and with little effect on the microfilaments that we believe accounts for the membrane protrusion. The CB treatment reduced the amount of microfilaments but had little effect on the microtubules. Most demecolcine-induced membrane protrusions disappeared when exposed to CB. Western blotting showed that CB treatment increases G-actin, which indicates a decrease in the microfilaments. High oocyte enucleation, survival, and developmental rates occurred when demecolcine-assisted enucleation was carried out in a CB-free solution. Higher blastocyst development rates and blastocyst cell numbers were achieved compared to control, indicating that CB is not necessary in the enucleation procedure of bovine oocytes. This study provides a clearer understanding of the mechanism for the demecolcine-induced oocyte membrane protrusion, and substantiates the practical use of demecolcine-assisted enucleation in a CB-free environment.
Topics: Actin Cytoskeleton; Animals; Cattle; Cell Membrane; Cell Nucleus; Cell Survival; Cytochalasin B; Demecolcine; Female; Microtubules; Nuclear Transfer Techniques; Oocytes; Tubulin Modulators
PubMed: 21740270
DOI: 10.1089/cell.2011.0012 -
Cell Structure and Function Dec 1994Demecolcine (Colcemid), an inhibitor of spindle fiber formation in M phase, induced apoptosis in V79 cells. At a concentration of 0.01 microgram/ml demecolcine, V79...
Demecolcine (Colcemid), an inhibitor of spindle fiber formation in M phase, induced apoptosis in V79 cells. At a concentration of 0.01 microgram/ml demecolcine, V79 cells proliferated exponentially as well as controls, although temporal M phase accumulation occurred 6 h after the addition of demecolcine. At 0.1 microgram/ml, the cells became hyperploid after remaining in the M phase for some time. Apoptosis occurred in V79 cells exposed to demecolcine at a concentration of 0.03 microgram/ml. Apoptosis was defined as the appearance of a sub-G1 peak in DNA histograms and a ladder pattern of fragmented DNA in gelelectrophoresis.
Topics: Animals; Apoptosis; Cell Cycle; Cell Line; Cricetinae; Cricetulus; DNA Damage; Demecolcine; Dose-Response Relationship, Drug; Lung; Ploidies
PubMed: 7720099
DOI: 10.1247/csf.19.391 -
Biology of Reproduction Apr 2003Studies were designed to further explore the use of pharmacological agents to produce developmentally competent enucleated mouse oocytes for animal cloning by somatic...
Demecolcine-induced oocyte enucleation for somatic cell cloning: coordination between cell-cycle egress, kinetics of cortical cytoskeletal interactions, and second polar body extrusion.
Studies were designed to further explore the use of pharmacological agents to produce developmentally competent enucleated mouse oocytes for animal cloning by somatic cell nuclear transfer. Metaphase II oocytes from CF-1 and B6D2F1 strains were activated with ethanol and subsequently exposed to demecolcine at various times postactivation. Chromosome segregation, spindle dynamics, and polar body (PB) extrusion were monitored by fluorescence microscopy using DNA-, microtubule-, and microfilament-selective probes. Exposure to demecolcine did not affect rates of oocyte activation induced by ethanol but did disrupt the coordination of cytokinesis and karyokinesis, suppressing the extent and completion of spindle rotation and second PB extrusion in a strain-dependent manner. Moreover, strain- and treatment-specific variations in the rate of oocyte enucleation were also detected. In particular, CF1 oocytes were more efficiently enucleated relative to B6D2F1 oocytes, and demecolcine treatments initiated early after activation resulted in higher enucleation rates than when treatment was delayed. The observed strain differences are possibly caused by a combination of factors, such as the time course of meiotic cell-cycle progression after ethanol activation, the degree of spindle rotation, and the extent of second PB extrusion. These results suggest that developmentally competent cytoplasts can be produced by timely exposure of activated oocytes to agents that disrupt spindle microtubules. However, the utility of the demecolcine-induced enucleation protocol will require further investigation into factors linking karyokinesis to cytokinesis at the levels of cell-cycle control and oocyte cytoskeletal remodeling following artificial or natural means of egg activation.
Topics: Animals; Animals, Outbred Strains; Cell Cycle; Cell Nucleus; Cloning, Organism; Cytoskeleton; Demecolcine; Ethanol; Female; Kinetics; Meiosis; Mice; Mice, Inbred Strains; Nuclear Transfer Techniques; Oocytes; Phenotype; Species Specificity
PubMed: 12606412
DOI: 10.1095/biolreprod.102.008292 -
The Journal of Reproduction and... Apr 2008The present study was carried out to examine whether demecolcine and sucrose affect the formation of a cytoplasmic protrusion containing chromosomes in pig oocytes...
The present study was carried out to examine whether demecolcine and sucrose affect the formation of a cytoplasmic protrusion containing chromosomes in pig oocytes independently or in combination. In the presence of 20 mM sucrose, the rates of oocytes with a cytoplasmic protrusion after culture for 60 min with 0.2-1.0 microg/ml demecolcine were significantly higher than those with 0.01-0.05 microg/ml demecolcine. When oocytes were cultured for 15 min in the presence of 0.2 microg/ml demecolcine and 20 mM sucrose, 35.1% of them extruded a cytoplasmic protrusion; this rate was significantly lower than those of oocytes cultured for 30-90 min. In the presence of 0.2 microg/ml demecolcine, significantly fewer oocytes extruded a cytoplasmic protrusion after culture for 30 min with 160 mM sucrose than with 0-80 mM sucrose. Significantly more oocytes extruded a cytoplasmic protrusion after culture for 30 min with 0.2 microg/ml demecolcine than without it, regardless of the presence or absence of 20 mM sucrose. In 88.9-100% of the oocytes, the cytoplasmic protrusions contained chromosomes with no significant differences among the different concentrations of demecolcine and sucrose and among the different treatment times. The results of the present study show that the cytoplasmic protrusion containing chromosomes in the pig oocyte is attributable to demecolcine, but sucrose does not affect its formation.
Topics: Animals; Chromosomes; Cloning, Organism; Cytoplasm; Demecolcine; Embryo Transfer; Female; Models, Biological; Nuclear Transfer Techniques; Oocytes; Sucrose; Swine; Time Factors; Tubulin Modulators
PubMed: 18239352
DOI: 10.1262/jrd.19142 -
Demecolcine toxicity: a case report of severe hematopoietic toxicity and a review of the literature.The American Journal of Medicine Sep 1959
Topics: Colchicine; Demecolcine; Hematopoiesis
PubMed: 13816950
DOI: 10.1016/0002-9343(59)90017-8 -
Reproduction in Domestic Animals =... Feb 2014The objective of our study was to establish the feasibility of experimental protocols for cloning sika deer. We performed auxiliary enucleation to improve the efficiency...
The objective of our study was to establish the feasibility of experimental protocols for cloning sika deer. We performed auxiliary enucleation to improve the efficiency of nuclear transfer operation by optimizing the demecolcine concentration to induce cytoplasmic protrusions in the sika deer oocytes. In the present study,we had studied the impact of different demecolcine concentrations on cytoplasmic protrusions and enucleation rates. We determined that 95.9% of the sika deer oocytes formed cytoplasmic protrusions when treated for 1 h with 0.8 μg/ml demecolcine. The lowest observed rate of protrusion was 19.3% after overnight treatment with demecolcine. When the oocytes aged or had a poor cumulus expansion, they exhibited a significant decrease in the ability to form cytoplasmic protrusions. The rates of enucleation (94.9% vs 85.8%, p < 0.05), cell fusion (84.6% vs 70.1%, p < 0.05) and blastocyst formation (15.4% vs 10.9%, p < 0.05) using demecolcine auxiliary enucleation were significantly higher than those after blind enucleation. These results demonstrated that sika deer oocytes could be enucleated quickly and effectively using demecolcine auxiliary enucleation, which could enhance the enucleation rate, cell fusion rate and blastocyst rate of cloned embryos in vitro.
Topics: Animals; Blastocyst; Cell Nucleus; Cloning, Organism; Cytoplasm; Deer; Demecolcine; Embryo Culture Techniques; Embryonic Development; Female; Nuclear Transfer Techniques; Oocytes; Tubulin Modulators
PubMed: 24138424
DOI: 10.1111/rda.12246 -
Cell Biology International Dec 2009To establish experimental protocols for cloning golden hamsters, optimal concentrations of colchicine and demecolcine were determined for inducing cytoplasmic protrusion...
To establish experimental protocols for cloning golden hamsters, optimal concentrations of colchicine and demecolcine were determined for inducing cytoplasmic protrusion (containing chromosomes) and assisting enucleation of their oocytes. Denuded oocytes at different ages were treated with 2.5-10 microg/ml of colchicine for 1-4h or 0.02-0.6 microg/ml of demecolcine for 15-60 min. Cytoplasmic protrusions of oocytes were removed with a micromanipulation pipette. The results show that: 1) at 13.5-18h post-hCG injection, approximately 90% of oocytes treated for with 10 microg/ml of colchicine formed cytoplasmic protrusions, and in some oocytes enucleation occurred; 2) when treated with 0.4 microg/ml of demecolcine for 1h, cytoplasmic protrusions 13.5-18h post-hCG treatment were present in almost all oocytes; 3) after the protrusions induced by either treatment had been removed, the assisted enucleation rate was >80%, whereas it was approximately 32% with blind enucleation.
Topics: Animals; Cell Nucleus; Cell Surface Extensions; Chorionic Gonadotropin; Chromosomes, Mammalian; Cloning, Organism; Colchicine; Cricetinae; Demecolcine; Female; Nuclear Transfer Techniques; Oocytes; Time Factors; Tubulin Modulators
PubMed: 19732847
DOI: 10.1016/j.cellbi.2009.08.010