New DNA probes to detect aneugenicity in rat bone marrow micronucleated cells by a pan-centromeric FISH analysis

Takeiri, Akira; Motoyama, Shigeki; Matsuzaki, Kaori; Harada, Asako; Taketo, Junko; Katoh, Chiaki; Tanaka, Kenji; Mishima, Masayuki

doi:10.1016/j.mrgentox.2013.05.011

Cited by 9 publications

(4 citation statements)

References 32 publications

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“…To characterize large micronucleus (LMN) in hepatocytes, FISH analysis for HEP specimens was conducted using a modification of a method that was previously reported for analysis of bone marrow erythrocyte specimens (Takeiri et al 2013). Template DNA derived from hepatocytes from male F344 rats, and primers (5′-TCC CGC TTG GAA CGA AGA GA-3′ and 5′-TTC TAT ATC CCG AAC AGT CC-3′) specific for centromeric satellite I (de Stoppelaar et al 1997;Takeiri et al 2013) were used for PCR amplification. PCR was conducted using Ex Taq (Takara Bio, Shiga, Japan) and a previously reported amplification protocol (Takeiri et al 2013).…”

Section: Fluorescence In Situ Hybridization (Fish) Analysis For Hep Specimensmentioning

confidence: 99%

“…Template DNA derived from hepatocytes from male F344 rats, and primers (5′-TCC CGC TTG GAA CGA AGA GA-3′ and 5′-TTC TAT ATC CCG AAC AGT CC-3′) specific for centromeric satellite I (de Stoppelaar et al 1997;Takeiri et al 2013) were used for PCR amplification. PCR was conducted using Ex Taq (Takara Bio, Shiga, Japan) and a previously reported amplification protocol (Takeiri et al 2013). PCR products were purified using NucleoSpin Gel and a PCR Clean-up kit (Macherey-Nagel, Düren, Germany).…”

Section: Fluorescence In Situ Hybridization (Fish) Analysis For Hep Specimensmentioning

confidence: 99%

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Chromosome aberrations induced by the non-mutagenic carcinogen acetamide involve in rat hepatocarcinogenesis through micronucleus formation in hepatocytes

et al. 2021

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Chromosome aberrations (CAs), i.e. changes in chromosome number or structure, are known to cause chromosome rearrangements and subsequently tumorigenesis. However, the involvement of CAs in chemical-induced carcinogenesis is unclear. In the current study, we aimed to clarify the possible involvement of CAs in chemical carcinogenesis using a rat model with the non-mutagenic hepatocarcinogen acetamide. In an in vivo micronucleus (MN) test, acetamide was revealed to induce CAs specifically in rat liver at carcinogenic doses. Acetamide also induced centromere-positive large MN (LMN) in hepatocytes. Immunohistochemical and electron microscopic analyses of the LMN, which can be histopathologically detected as basophilic cytoplasmic inclusion, revealed abnormal expression of nuclear envelope proteins, increased heterochromatinization, and massive DNA damage. These molecular pathological features in LMN progressed with acetamide exposure in a timedependent manner, implying that LMN formation can lead to chromosome rearrangements. Overall, these data suggested that CAs induced by acetamide play a pivotal role in acetamide-induced hepatocarcinogenesis in rats and that CAs can cause chemical carcinogenesis in animals via MN formation.

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Section: Fluorescence In Situ Hybridization (Fish) Analysis For Hep Specimensmentioning

confidence: 99%

Section: Fluorescence In Situ Hybridization (Fish) Analysis For Hep Specimensmentioning

confidence: 99%

Chromosome aberrations induced by the non-mutagenic carcinogen acetamide involve in rat hepatocarcinogenesis through micronucleus formation in hepatocytes

et al. 2021

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“…This allowed a development strategy for the screening stage to be established without conducting any further mechanistic studies. An Ames test, an in vitro MNT accompanied by centromeric FISH analysis, and a rat MNT with FISH analysis [ 22 ] were chosen as the studies for regulatory application. If γH2AX had not been evaluated at the early stage, the candidates would have been developed without ruling out the risk that the compounds were clastogenic.…”

Section: Use Of γH2ax Combined With Other Endpoints In Early Screeninmentioning

confidence: 99%

Advantages of evaluating γH2AX induction in non-clinical drug development

Motoyama¹,

Takeiri²,

Tanaka³

et al. 2018

Genes and Environ

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γH2AX, the phosphorylated form of a histone variant H2AX at Ser 139, is already widely used as a biomarker to research the fundamental biology of DNA damage and repair and to assess the risk of environmental chemicals, pollutants, radiation, and so on. It is also beginning to be used in the early non-clinical stage of pharmaceutical drug development as an in vitro tool for screening and for mechanistic studies on genotoxicity. Here, we review the available information on γH2AX-based test systems that can be used to develop drugs and present our own experience of practically applying these systems during the non-clinical phase of drug development. Furthermore, the potential application of γH2AX as a tool for in vivo non-clinical safety studies is also discussed.

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“…Our model predicts that CENP-C has co-evolved with interacting partners to modulate effector recruitment, so that an allele from another species will disrupt these interactions and weaken the kinetochore pathway (Figure 1H, Prediction 1). To test this prediction, we selected rat as a model organism close to mouse with divergent centromere DNA and proteins (Gibbs et al, 2004; Takeiri et al, 2013). Because protein interfaces change by genetic drift as well as by selection, an allele from a closely related species minimizes incompatibilities coming from stochastic changes.…”

Section: Introductionmentioning

confidence: 99%

Centromere drive and suppression by parallel pathways for recruiting microtubule destabilizers

Kumon

Stefanik

et al. 2020

Preprint

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SummarySelfish centromere DNA sequences bias their transmission to the egg in female meiosis. Evolutionary theory suggests that centromere proteins evolve to suppress costs of this “centromere drive”. In hybrid mouse models with genetically different maternal and paternal centromeres, selfish centromere DNA exploits a kinetochore pathway to recruit microtubule-destabilizing proteins that act as drive effectors. We show that such functional differences are suppressed by a parallel pathway for effector recruitment by heterochromatin, which is similar between centromeres in this system. Disrupting heterochromatin by CENP-B deletion amplifies functional differences between centromeres, whereas disrupting the kinetochore pathway with a divergent allele of CENP-C reduces the differences. Molecular evolution analyses using newly sequenced Murinae genomes identify adaptive evolution in proteins in both pathways. We propose that centromere proteins have recurrently evolved to minimize the kinetochore pathway, which is exploited by selfish DNA, relative to the heterochromatin pathway that equalizes centromeres, while maintaining essential functions.

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New DNA probes to detect aneugenicity in rat bone marrow micronucleated cells by a pan-centromeric FISH analysis

Cited by 9 publications

References 32 publications

Chromosome aberrations induced by the non-mutagenic carcinogen acetamide involve in rat hepatocarcinogenesis through micronucleus formation in hepatocytes

Chromosome aberrations induced by the non-mutagenic carcinogen acetamide involve in rat hepatocarcinogenesis through micronucleus formation in hepatocytes

Advantages of evaluating γH2AX induction in non-clinical drug development

Centromere drive and suppression by parallel pathways for recruiting microtubule destabilizers

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