In vivo assays for detection and evaluation of aneugens
Conference
·
OSTI ID:28455
- State Univ. of Leiden (Netherlands)
- Medscand Ingeny B.V., Leiden (Netherlands)
Three different assays were developed and standardized to study the aneugenic potential of known or suspected aneugens in vivo. (1) The cytokinesis blocked micronuclei (MN) assay and fluorescent in-situ hybridization technique using mouse centromeric satellite DNA were applied to study aneuploidy in Swiss random bred mice following treatment with Hexamethyl-phosphoramide (HMPA), diethylstilbestrol (DES) and whole body X-irradiation. The MN in the binucleated splenocytes were scored as centromere specific DNA positive (C+) or centromere specific DNA negative (C-) depending on the presence of the fluorescent signal in the MN following in-situ hybridization with centromere specific DNA. Both chemicals induced significant number of C+ micronuclei (in the range of 60-89%) in cytokinesis blocked mouse splenocytes. X-irradiation induced MN frequency in a dose dependent manner and approximately 23% were C+. (2) Transgenic mice with multiple copies of foreign DNA (C myc and lambda) inserted at a single site lambda on chromosome 2 and X, C myc on chromosome (8) were generated and used for detecting aneugens on the basis that the chromosome carrying specific foreign DNA inserts which can be detected easily in both metaphase and interphase cell and different organs using DNA probes and in-situ hybridisation. Following treatment of transgenic mice with DES and vinblastine sulfate frequency of aneuploidy increased significantly in bone marrow and spleen cells. However, induction of aneuploidy was not correlated with dose of the chemicals. (3) DNA libraries constructed for specific mouse chromosomes, biotin labelled and used as probe for in situ hybridisation to study aneuploidy in vivo.
- OSTI ID:
- 28455
- Report Number(s):
- CONF-9210475--Cond.
- Country of Publication:
- United States
- Language:
- English
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