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Micronuclei and nuclear buds in amniotic tissue of rats treated with cyclophosphamide
Ramón Guillermo Ortiz-García1, Belinda Claudia Gómez-Meda2, Juan Ernesto Gutiérrez-Sevilla3
1Doctorado en Genética Humana, Departamento de Biología Molecular y Genómica, Centro Universitario de Ciencias de la Salud, Universidad de Guadalajara, Guadalajara, Jalisco, Mexico; Laboratorio de Mutagénesis, División de Medicina Molecular, Centro de Investigación Biomédica de Occidente, Instituto Mexicano del Seguro Social, Guadalajara, Jalisco, Mexico.
Abstract:
Fetal development can be altered by DNA damage caused by maternal exposure to chemical, physical, or biological agents during gestation. One method of assessing genotoxicity is to detect micronuclei (MNs) and/or nuclear abnormalities. This can be performed in vivo and requires only frequently dividing tissues, such as amniotic tissue (AT), which is in contact with the fetal environment and is composed of very thin layers of cells. This study evaluated the presence of MNs, nucleoplasmic bridges, and nuclear buds (NBs) in the fetal AT following maternal exposure to cyclophosphamide (CP) during pregnancy. Pregnant Wistar rats were divided into a negative control group and an experimental group that was orally administered CP (10 mg/kg). Daily blood smears were obtained from pregnant rats on days 14-19 of gestation. The rats were dissected, and fetal ATs were obtained on the 19th day of gestation. The MN and NB frequencies in AT cells were analyzed using a fluorescence microscope (100 ×). Micronucleated erythrocytes in the peripheral blood of the control rats were also assessed. Micronucleated polychromatic erythrocyte frequencies were significantly higher than those in the controls. Polychromatic erythrocyte frequencies were lower in CP-treated rats than in controls at 48-120 h. Fetuses in the CP-treated group also showed a significant increase in MNs and NBs in AT cells. In conclusion, AT could be used for analyzing MNs and NBs in rats following maternal exposure to a genotoxic agent and as a viable alternative for analyzing the integrity of fetal DNA during gestation.
Insights
Maternal exposure to genotoxic agents during pregnancy can harm fetal development. Amniotic tissue (AT) effectively detects DNA damage, such as micronuclei (MNs) and nuclear buds (NBs), in fetuses following maternal exposure.
Area of Science:
- Toxicology and Environmental Health
- Developmental Biology
- Genetics and Genomics
Background:
- Maternal exposure to genotoxic agents during gestation can adversely affect fetal development.
- Assessing fetal genotoxicity is crucial for understanding developmental risks.
- Micronuclei (MNs) and nuclear abnormalities are key biomarkers of DNA damage.
Purpose of the Study:
- To evaluate the presence of MNs, nucleoplasmic bridges, and nuclear buds (NBs) in fetal amniotic tissue (AT) after maternal exposure to cyclophosphamide (CP).
- To determine if fetal AT is a suitable tissue for in vivo genotoxicity assessment.
- To assess the integrity of fetal DNA following maternal genotoxic insult.
Main Methods:
- Pregnant Wistar rats were administered cyclophosphamide (CP) orally (10 mg/kg) or served as controls.
- Fetal amniotic tissues (AT) were collected on day 19 of gestation.
- Micronuclei (MNs) and nuclear buds (NBs) in AT cells were analyzed using fluorescence microscopy; peripheral blood micronucleated erythrocytes were also assessed.
Main Results:
- CP-treated rats showed a significant increase in MNs and NBs in fetal AT cells compared to controls.
- Micronucleated polychromatic erythrocyte frequencies were significantly higher in CP-treated rats.
- Polychromatic erythrocyte frequencies decreased in CP-treated rats, indicating potential bone marrow toxicity.
Conclusions:
- Fetal amniotic tissue (AT) is a viable and sensitive option for detecting DNA damage (MNs and NBs) in fetuses exposed to genotoxic agents in utero.
- This study validates AT as an alternative method for assessing fetal DNA integrity during gestation.
- Findings highlight the importance of monitoring maternal exposure to genotoxic substances during pregnancy.
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