Detection of chemically induced DNA lesions in multiple mouse organs (liver, lung, spleen, kidney, and bone marrow) using the alkaline single cell gel electrophoresis (Comet) assay

Sasaki, Yu; Tsuda, Shuji; Izumiyama, Fusako; Nishidate, Emi

doi:10.1016/s1383-5718(96)00133-7

Cited by 143 publications

(70 citation statements)

References 15 publications

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“…To obtain nuclei, the homogenate was centrifuged at 700 g for 10 min at 08C, and the precipitate was resuspended in chilled homogenizing buffer at 0.5 g=mL and allowed to settle; precipitated clumps were then removed. Doses and times were selected based on the preliminary studies as well as literature-reported values (Sasaki et al, 1997a(Sasaki et al, , 1997bTsuda et al, 2000;Sekihashi et al, 2002).…”

Section: Treatment and Organ Preparationmentioning

confidence: 99%

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Hosseinzadeh

Abootorabi

Sadeghnia

2008

DNA and Cell Biology

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This study was designed to examine the effect of aqueous extract of Crocus sativus stigmas (CSE) and crocin (trans-crocin 4) on methyl methanesulfonate (MMS)-induced DNA damage in multiple mice organs using comet assay. Adult male NMRI mice in different groups were treated with either physiological saline (10 mL=Kg, intraperitoneal [ip]), CSE (80 mg=Kg, ip), crocin (400 mg=Kg, ip), MMS (120 mg=Kg, ip), and CSE (5, 20, and 80 mg=Kg, ip) 45 min prior to MMS administration or crocin (50, 200, and 400 mg=Kg, ip) 45 min prior to MMS administration. Mice were scarified about 3 h after each different treatment, and the alkaline comet assay was used to evaluate the effect of these compounds on DNA damage in different mice organs. The percent of DNA in the comet tail (% tail DNA) was measured. A significant increase in the % tail DNA was seen in nuclei of different organs of MMS-treated mice. In control groups, no significant difference was found in the % tail DNA between CSE-or crocin-pretreated and saline-pretreated mice. The MMS-induced DNA damage in CSEpretreated mice (80 mg=Kg) was decreased between 2.67-fold (kidney) and 4.48-fold (lung) compared to those of MMS-treated animals alone ( p < 0.001). This suppression of DNA damage by CSE was found to be depended on the dose, which pretreatment with CSE (5 mg=Kg) only reduced DNA damage by 6.97%, 6.57%, 7.27%, and 9.90% in liver, lung, kidney, and spleen, respectively ( p > 0.05 as compared with MMS-treated group). In the same way, crocin also significantly decreased DNA damage by MMS (between 4.69-fold for liver and 6.55-fold for spleen, 400 mg=Kg), in a dose-dependent manner. These data indicate that there is a genoprotective property in CSE and crocin, as revealed by the comet assay, in vivo.

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Section: Treatment and Organ Preparationmentioning

confidence: 99%

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Hosseinzadeh

Abootorabi

Sadeghnia

2008

DNA and Cell Biology

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“…This experiment followed the design of our previous comet assay studies on multiple mouse organs (16)(17)(18). From shortly after they were treated until just before they were killed, the animals were carefully observed for pharmacotoxic signs and they were killed by bleeding under anesthesia 3 or 24 h after the treatment, and then 8 organs-glandular stomach, colon, liver, kidney, urinary bladder, lung, brain, and bone marrowwere removed.…”

Section: Methodsmentioning

confidence: 99%

In vivo Genotoxic Potential of Kojic Acid in Rodent Multiple Organs Detected by the Comet Assay

Kawaguchi

Nakamura

Honda

et al. 2008

Genes and Environment

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Kojic acid has been used for skin whitening as a cosmetic agent. Kojic acid is believed to be hepatocarcinogenic in mice. We conducted the comet assay in mouse and rat multiple organs to evaluate its in vivo genotoxic potential. Kojic acid induced dose-dependent DNA damage in ddY mouse stomach and liver and in Wistar rat stomach, liver, lung, and bone marrow after a single gavage administration at ≦1000 mg/kg. Its hepatic genotoxicity detected by the comet assay seems to contradict to absence of its hepatic tumor initiating activity revealed by the two-step carcinogenesis studies with mice and rats. However, considering the possibility that the sensitivity of the two-step carcinogenesis studies performed are not high enough to detect weak initiating activity of a chemical, it would be premature to conclude that its hepatic genotoxicity contradicts to the absence of initiating activity. In mice fed a diet containing 3% kojic acid for up to 10 days, DNA migration increased in the stomach after feeding for 2 days but it did not increased after feeding for 1 day and ≧4 days. In the colon, DNA migration after feeding of 3% kojic acid for 2 days was higher than the control values, but this increase was not statistically signiˆcant. In the stomach and colon, any statistically signiˆcant increases in DNA migration were not observed after feeding of 1.5% kojic acid. In the liver, DNA migration increased with feeding period and the increases after feeding of 3% and 1.5% kojic acid for ≧4 days and 6 days, respectively, were statistically signiˆcant. Our present results suggested good correlation between hepatocarcinogenicity of kojic acid and increasing tendency of its hepatic genotoxicity with dosing period when it is given to mice continuously in the diet.

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“…The former is very important so as to avoid pseudonegative results and the comet assay can detecta wide variety of genotoxic compounds both in vitro [35,36] and in vivo [3,[37][38][39][40]. The micronucleus test (MN test) is a standard procedure that can detect structural chromosome aberrations derived from initial damage in the S phase and/or numerical chromosome aberrations due to an eugenic effects in the M phase [40,41].…”

Section: Power Of the Comet Assay To Detect Low Level Genotoxicitymentioning

confidence: 99%

The Power of the Comet Assay to Detect Low Level Genotoxicity and DNA Repair Factors Affecting its Power

Sasaki

SatomiKawaguchi

Nakamura

2017

MOJT

Self Cite

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The comet assay is considered to be a rapid and sensitive procedure for quantitating DNA damage in mammalian cells. In this article, to interpret its outcomes adequately, its power to detect low level genotoxicity and factors affecting its power were reviewed. Although the development of initial lesions into alkali-labile sites and/or SSBs through repairing events is an important factor to support its detecting power, their repair reduce its power to detect SSBs as an initial DNA lesion. The acellular comet assay, in which slides with gel are prepared from untreated cells are exposed after lysis to test agents. Thus, detection of SSBs as initial lesions but not alkali-labile sites generated from DNA lesion such as alkylated bases and the power to detect low level SSBs as initial lesions is lower in the standard than in the acellular assay. The acellular comet assay would be practically used to detect SSBs as initial lesions. The inhibitors of re-synthesis andincisionsteps of the excision repair enhance and suppress comet-tail formation, respectively. Although the incision step of repairing process is necessary to support the sensitivity of the comet assay, its re-synthesis step reduces the power to detect low level genotoxicity. Although this assay can detect a wide variety of genotoxoic compounds and its power to detect genotoxicity were identical to that of the MN test, its power to detect low levels of genotoxic potential is inferior to that of the MN test. However, its power of to detect a low level of genotoxicity can be elevated to a level higher than that of the MN test by using DNA resynthesis inhibitors, such as araC and HU. In conclusion, the outcomes of comet assay should be interpretedappropriately in consideration of the action of DNA repair.

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Detection of chemically induced DNA lesions in multiple mouse organs (liver, lung, spleen, kidney, and bone marrow) using the alkaline single cell gel electrophoresis (Comet) assay

Cited by 143 publications

References 15 publications

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

Protective Effect of Crocus sativus Stigma Extract and Crocin (trans-crocin 4) on Methyl Methanesulfonate–Induced DNA Damage in Mice Organs

In vivo Genotoxic Potential of Kojic Acid in Rodent Multiple Organs Detected by the Comet Assay

The Power of the Comet Assay to Detect Low Level Genotoxicity and DNA Repair Factors Affecting its Power

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