Mutant frequencies and spectra depend on growth state and passage number in cells cultured from transgenic lacZ-plasmid reporter mice.
Name:
Publisher version
View Source
Access full-text PDFOpen Access
View Source
Check access options
Check access options
Average rating
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Star rating
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Type
ArticleLanguage
en
Metadata
Show full item recordTitle
Mutant frequencies and spectra depend on growth state and passage number in cells cultured from transgenic lacZ-plasmid reporter mice.Publiekssamenvatting
Transgenic mice harboring the lacZ gene within a plasmid that can be recovered and amplified in Escherichia coli, to establish mutant frequencies and spectra, have provided crucial insights into the relationships between mutations, cancer and aging in vivo. Here, we use embryonic fibroblasts from transgenic lacZ-plasmid reporter mice to determine the relationship between cell proliferation in culture and mutations induced by ultraviolet (UV) light. A single dose of 2.5J/m2 of UVC to actively proliferating cells caused an approximately eight-fold increase in mutant frequency 24 h after irradiation. Identically treated quiescent cells showed a two-fold increase in mutant frequency. Thus, whereas proliferation facilitated the acquisition of mutations, it was not an absolute requirement. Characterization of the UV-induced mutations indicated that the lower mutant frequency in quiescent cells was due mainly to a reduction in point mutations; size-change mutations, indicative of translocations or deletions, were relatively unaffected by the growth state of the cells. To investigate long-term genomic stability after UVC-induced damage, we monitored the lacZ locus in irradiated cells passaged for many generations in culture. The results indicated the emergence of jackpot mutations of rapidly changing frequency, most likely reflecting the successive emergence and decline of dominant cell clones during long-term culture. These findings show that the lacZ-plasmid locus is a valid reporter for studying induced mutations in short-term cultures of both quiescent and proliferating fibroblasts. In long-term cultures, the locus is less suitable for studying induced mutations owing to the instability of the cell population.PMID
16126462ae974a485f413a2113503eed53cd6c53
10.1016/j.dnarep.2005.07.006
Scopus Count
Collections
Related articles
- Background mutations and polymorphisms in lacZ-plasmid transgenic mice.
- Authors: Dollé ME, Snyder WK, van Orsouw NJ, Vijg J
- Issue date: 1999
- Treatment of lacZ plasmid-based transgenic mice with benzo[a]pyrene: measurement of DNA adduct levels, mutant frequencies, and mutant spectra.
- Authors: Boerrigter ME
- Issue date: 1999
- High sensitivity for color mutants in lacZ plasmid-based transgenic mice, as detected by positive selection.
- Authors: Boerrigter ME
- Issue date: 1998
- Mutation spectrum in UVB-exposed skin epidermis of Xpa-knockout mice: frequent recovery of triplet mutations.
- Authors: Ikehata H, Yanase F, Mori T, Nikaido O, Tanaka K, Ono T
- Issue date: 2007 Jan
- Sources of variability in mutant frequency determinations in different organs of lacZ plasmid-based transgenic mice: experimental features and statistical analysis.
- Authors: Boerrigter ME, Vijg J
- Issue date: 1997