Dominant cataract mutations and specific-locus mutations in mice induced by radiation or ethylnitrosourea

Ehling, U.H.; Favor, Jack; Kratochvílová, Jana; Neuhäuser-Klaus, A.

doi:10.1016/0027-5107(82)90222-6

Cited by 108 publications

(22 citation statements)

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“…Exposures to identical doses of ENU have yielded germcell mutation inductions from =50-to 200-fold above background when the visible specific-locus test is used (15,26), and induced mutant frequencies can vary from =10-to >140-fold above background when individual loci are examined separately (26). Few ENU-induced mutations detected in the visible specific-locus test have been characterized; whether the presence of hotspots for ENU within the seven loci plays a role in these high inductions is unknown.…”

Section: Discussionmentioning

confidence: 99%

Characterization of mutations induced by ethylnitrosourea in seminiferous tubule germ cells of transgenic B6C3F1 mice.

Provost¹,

Short²

1994

Proc. Natl. Acad. Sci. U.S.A.

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Transgenic B6C3F1 mice carrying a lacl target gene were exposed to acute and multiple doses of ethylnitrosourea (ENU), and germ cells from the seminiferous tubules were assayed for mutation 3 and 90 days after treatment. Relative to untreated controls, the mutation frequency increased 3.2-and 19.9-fold at 3 and 90 days after treatment, respectively. Mutant lacl genes recovered from untreated and treated groups were sequenced, and the spectra of mutations were determined. Eighty-flve percent (11/13) of the spontaneous mutatious resulted in G-C -* AT transitions, all ofwhich occurred at CpG dinucleotides. Fifteen of 22 sites (68%) found mutated 3 days after ENU treatment occurred at G-C base pairs, although some of these are expected to be spontaneous mutations. Ninety days after treatment, 13 of 19 sites (68%) found mutated occurred at APT base pairs. The mutation spectra seen are consistent with proposed mechanisms of ENU mutagenesis and correlate with the in viva spectra seen in ENU studies by using transmissibiliy assays and the hpilgene. These findings represent significant progress toward dening the in vivo spectra of Ethylnitrosourea (ENU) induces a broad spectrum of promutagenic lesions in vivo (6, 7) and has been studied by using a variety of transmissibility assays (8)(9)(10)(11)(12)(13)(14)(15). Although ENU has been well characterized as a mouse germ cell mutagen, the spectrum of ENU-induced mutations in mouse germ cells remains largely undefined. To date, only six germ-line mutations identified in ENU-treated mice offspring have been characterized at the nucleotide level and reported, all of which occur at either adenine or thymine residues (10-13, 16). The experiments described here characterize the mutagenic response oftransgenic mouse testicular germ cells to acute-and multiple-dose ENU exposure and outline the spectra of mutants recovered after these exposures. These findings represent progress toward defining the in vivo spectrum of ENU mutagenesis in germ cells and explore the relationship to the in vivo somatic cell spectrum in the endogenous hprt gene. MATERIALS AND METHODSTransgenic Mice. Transgenic B6C3F1 hybrid mice were generated from natural matings (performed at Taconic Farms) of inbred female transgenic hemizygous A/lacd C57BL/6 mice (2) and male nontransgenic C3H mice. DNA was prepared from tail tips of the resulting offspring and screened for the transgene by dot-blot DNA hybridization, as described (2). Animals were maintained on a 12-hr light schedule in cages containing heat-sterilized wood chips and were provided with F4 rodent block food (Harlan Bioproducts, Haslett, MI) and tap water ad libitum.Mutagen Treatment. ENU (Sigma, Lot no. 20H0369) was dissolved in Dulbecco's phosphate-buffered saline (pH 6.0) (GIBCO) and administered within 30 min of solution preparation. Four 8-to 10-week-old male transgenic B6C3F1 mice (Big Blue mouse, Stratagene) were given three i.p. injections of 100 mg (± 2%) of ENU per kg of body weight at 7-day intervals for a cumulative multiple dos...

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Section: Discussionmentioning

confidence: 99%

Characterization of mutations induced by ethylnitrosourea in seminiferous tubule germ cells of transgenic B6C3F1 mice.

Provost¹,

Short²

1994

Proc. Natl. Acad. Sci. U.S.A.

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“…Additionally, hybrid B6C3F 1 transgenic male mice were purchased from Taconic Farms. T stock female mice [Searle, 1974;Russell and Matter, 1980;Ehling et al, 1982;Peters and Lewis, 1991], homozygous for seven recessive visible markers (non-agouti (a/a), brown (b/b), pink-eyed dilution (p/p), chinchilla (c ch /c ch ), dilute (d/d), short ear (se/se), and piebald (s/s)) were used for the morphological specific-locus test (MSLT). The T stock animals were originally obtained from the MRC Radiobiology Unit, Harwell (Dr. J. Peters) and have since been maintained as a random-bred stock at the RTI International Animal Research Facility for over a decade.…”

Section: Animals and Husbandrymentioning

confidence: 99%

Transmission of mutations in the lacI transgene to the offspring of ENU‐treated Big Blue® male mice

Barnett

Tyl

Shane

et al. 2002

Environ and Mol Mutagen

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The purpose of the study was to determine: 1) if male germ cells of Big Blue mice carrying newly induced mutations in the lacI transgene were effective in fertilization; 2) if offspring arising from such mutant sperm had the mutation in germ cells and multiple somatic tissues; and 3) how the frequency of mutants induced in the lacI transgene compared to the frequency induced in endogenous genes traditionally employed to study germ cell mutagenesis in mice. Male B6C3F(1) mice hemizygous for the lambda/lacI transgene were treated weekly with 100 mg/kg body weight of the mammalian germ cell mutagen N-ethyl-N-nitrosourea (ENU). The cumulative dose for each treated animal was 300 mg ENU/kg body weight. Ten weeks later the treated mice were mated to T stock females and the resulting offspring were screened for specific-locus mutations at six loci affecting external appearance, as well as for mutations in the lacI transgene in multiple somatic tissues and germ cells. Five offspring carrying recessive specific-locus mutations were observed among 597 offspring screened (mutant frequency = 139.6 x 10(-5) per locus). Four offspring carrying lacI mutations were observed among 280 offspring screened (mutant frequency = 35.7 x 10(-5) per locus (assuming 40 target loci)). Each of the four lacI mutant offspring carried a different mutation. Three of the mutations were A:T-->G:C transitions and one a G:C-->A:T transition. Consistent with the expectation that a mutation induced in a parental germ cell and transmitted to a conceptus would exist in every cell of the offspring, each mutant mouse had identical mutations in all somatic tissues sampled, as well as in its germ cells. These data provide preliminary evidence for the biological validity of assessing induced, heritable mutations using transgenic mice, without the need for generating an F(1) generation.

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“…A single wild type PAX6/Pax6 allele is not sufficient for normal eye development in human, mouse and rat implicating haploinsufficiency. Rodents harboring mutated Pax6 alleles are comprised of: spontaneously deleted alleles, including mouse Sey Dey and Sey H (for a review, see Glaser et al, 1995) and rat rSey (Matsuo et al, 1993), chemically-induced mutations, including mouse Pax6 1 to 10 Neu (Ehling et al, 1982;Favor et al, 2001), engineered disruption of Pax6 locus in mouse embryonic stem cells combined with insertion of a lacZ (St-Onge et al, 1996) and floxed Pax6 allele used for conditional inactivation of Pax6 (Ashery-Padan et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Regulation of gene expression by Pax6 in ocular cells: a case of tissue-preferred expression of crystallins in lens

Cvekl¹,

Yang²,

Chauhan³

et al. 2004

Int. J. Dev. Biol.

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Dominant cataract mutations and specific-locus mutations in mice induced by radiation or ethylnitrosourea

Cited by 108 publications

References 10 publications

Characterization of mutations induced by ethylnitrosourea in seminiferous tubule germ cells of transgenic B6C3F1 mice.

Characterization of mutations induced by ethylnitrosourea in seminiferous tubule germ cells of transgenic B6C3F1 mice.

Transmission of mutations in the lacI transgene to the offspring of ENU‐treated Big Blue® male mice

Regulation of gene expression by Pax6 in ocular cells: a case of tissue-preferred expression of crystallins in lens

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