Using fluorescence for improvement of the quantitative analysis of micronucleus in cell culture

Dias, Viviane Miranda; Manelli-Oliveira, Renata; Machado-Santelli, Gláucia Maria

doi:10.1016/j.mrgentox.2004.10.003

Cited by 22 publications

(16 citation statements)

References 28 publications

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“…Interphase cytogenetics is especially useful in genotoxicity studies as most mutagens decrease mitotic activity. Recently, the MN test with fluorescent stains proved to be more sensitive in detecting small micronuclei compared to the MN test using traditional methods, such as Feulgen’s reaction (Dias et al 2005). Therefore, the use of fluorescence has been proposed as an accurate method for the detection of micronuclei.…”

Section: Introductionmentioning

confidence: 99%

FISH in analysis of gamma ray-induced micronuclei formation in barley

Juchimiuk-Kwasniewska

Brodziak

Małuszyńska

2010

J Appl Genetics

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A micronucleus test in combination with fluorescent in situ hybridization (FISH) using telomere-, centromere-specific probes and 5S and 25S rDNA was used for a detailed analysis of the effects of gamma ray irradiation on the root tip meristem cells of barley, Hordeum vulgare (2n = 14). FISH with four DNA probes was used to examine the involvement of specific chromosomes or chromosome fragments in gamma ray-induced micronuclei formation and then to explain their origin. Additionally, a comparison of the possible origin of the micronuclei induced by physical and chemical treatment: maleic hydrazide (MH) and N-nitroso-N-methylurea (MNU) was done. The micronuclei induced by gamma ray could originate from acentric fragments after chromosome breakage or from whole lagging chromosomes as a result of a dysfunction of the mitotic apparatus. No micronuclei containing only centromeric signals were found. An application of rDNA as probes allowed it to be stated that 5S rDNA–bearing chromosomes are involved in micronuclei formation more often than NOR chromosomes. This work allowed the origin of physically- and chemically-induced micronuclei in barley cells to be compared: the origin of micronuclei was most often from terminal fragments. FISH confirmed its usefulness in the characterization of micronuclei content, as well as in understanding and comparing the mechanisms of the actions of mutagens applied in plant genotoxicity.

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Section: Introductionmentioning

confidence: 99%

FISH in analysis of gamma ray-induced micronuclei formation in barley

Juchimiuk-Kwasniewska

Brodziak

Małuszyńska

2010

J Appl Genetics

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“…Other studies also point to the role of nanoparticles in mediated DNA damages owing to the development of oxidative stress [15][16][17]. To estimate changes in the stability of cell genomes in systems in vivo and in vitro through the action of nano composites and nanoparticles, various methodological approaches are used, one of which is the MN test [5,10,11,18]. Modern researchers offer different ways of FM action, depending on their size and stabi lizer.…”

Section: Resultsmentioning

confidence: 99%

“…The number of micronuclei (MN) was evaluated on preparations col ored with acridine orange (3.0 mg/mL in a phosphate buffer with pH 6.4-6.5) for 1000 AEC cells with the help of a Micros MS 300 fluorescent microscope (40 × 10) [11].…”

Section: Methodsmentioning

confidence: 99%

“…As literature data show, a classical method for determining the genotoxic effects of harmful factors on live organisms is the micronucleum test and trans membrane mitochondrial potential of mitochondria [10][11][12]. The purpose of the present work was to evaluate the resistance of the genome and transmembrane mito chondrial potential of tumor cells to the impact of nanoferomahnetcs with different sizes of nanoparti cles in an in vivo system using two methodic approaches, namely, the micronucleum test and fluo rescent probe JC 1.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of the genome stability and mitochondrial potential of tumor cells in vivo under the action of nanoferromagnetics with different sizes of nanoparticles

et al. 2013

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The transmembrane potential and genome stability of tumor cells under nanoferromagnetic action with different sizes of nanoparticles were studied in vivo. An evaluation of the genotoxic influence of nanoeferromagnetics on tumor cells and changes in their mitochondrial potential was carried out with the application of a micronuclei test and mitochondrial fluorescent probe JC 1. Nanoferromagnetics with nano particle diameters of 40 and 100 nm at a concentration of 3 mg/kg were not found to deminish the trasmem brane mitochondrial potential and the genome of tumor cells and increase the micronucleus frequency of tumor cells to only a small extent. Iron nanoparticles at a concentration of 6 mg/kg were noted to demon strate an antidromonous character of the changing mitochondrial potential and micronuclei frequency depending on their size. Iron nanoparticles with a smaller diameter were found to provoke an essential rise in the transmembrane potential of tumor cell metachondria on the background of a constant number of micro nuclei, and the opposite was true for those with a greater diameter. The data obtained can serve as an impor tant criterion for further studies in experimental oncology to create an optimal system for delivering antican cer preparations to tissue targets.

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“…The mean frequency of MNIE in concurrent positive controls is higher in the first experiment (69.5 ± 11.7 MNIE/1000 IEs) compared to the second one (46.5 ± 7.7 MNIE/1000 IEs), evidencing the difference between the two different scoring methods applied in the main experiment and in the second one. The use of propidium iodide, counterstain for the DNA in the second experiment, presents advantages compared to acridine orange: the fluorescence is long lasting and longer than AO (Dias et al, 2005) • Individual data for MNIE/1000 cells (24.0, 33.5, 6.5, 8.0, 29.0) for buprofezin 2000 mg/kg in the first experiment, reveal a large interindividual variability.…”

mentioning

confidence: 99%

Opinion on Genotoxic and Carcinogenic Potential of Buprofezin in the Context of the Human Risk Assessment - Scientific Opinion of the Panel on Plant Protection Products and their Residues (PPR)

2008

EFSA Journal

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Using fluorescence for improvement of the quantitative analysis of micronucleus in cell culture

Cited by 22 publications

References 28 publications

FISH in analysis of gamma ray-induced micronuclei formation in barley

FISH in analysis of gamma ray-induced micronuclei formation in barley

Evaluation of the genome stability and mitochondrial potential of tumor cells in vivo under the action of nanoferromagnetics with different sizes of nanoparticles

Opinion on Genotoxic and Carcinogenic Potential of Buprofezin in the Context of the Human Risk Assessment - Scientific Opinion of the Panel on Plant Protection Products and their Residues (PPR)

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