Mitotic recombination produces the majority of recessive fibroblast variants in heterozygous mice.
Mice heterozygous at Aprt (adenine phosphoribosyltransferase) were used as a model to study in vivo loss of heterozygosity (LOH) in normal fibroblasts. Somatic cell variants that exhibited functional loss of the wild-type Aprt in vivo were recovered as APRT-deficient cell colonies after culturing in selection medium containing 2, 6-diaminopurine (DAP), an adenine analog that is toxic only to cells with APRT enzyme activity. DAP-resistant (DAP(r)) fibroblast variants were recovered at a median frequency of 12 x 10(-5) from individual ears from progeny of crosses between mouse strains 129/Sv and C3H/HeJ. The frequency of DAP(r) variants varied greatly among individual ears, suggesting that they preexisted in vivo and arose at various times during development. Polymorphic molecular markers and a cytological marker on the centromere of chromosome 8 made it possible to discriminate between each of six possible mechanistic pathways of LOH. The majority (about 80%) of the DAP(r) variants were a consequence of mitotic recombination. The prevalence of mitotic recombination in regions proximal to Aprt did not correlate with meiotic map distances. In particular, there was a higher than expected frequency of crossovers within the interval 59 cM to 67 cM. The high spontaneous frequency of Aprt LOH, mediated primarily by mitotic recombination, is fully consistent with our previous results with human peripheral T cells from individuals known to be heterozygous at APRT. Thus, this Aprt heterozygote mouse is a valid model for studying somatic mutagenesis and mitotic recombination in vivo.
Shao
C
C
Department of Medical and Molecular Genetics and The Walther Oncology Center, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Deng
L
L
Henegariu
O
O
Liang
L
L
Raikwar
N
N
Sahota
A
A
Stambrook
P J
PJ
Tischfield
J A
JA
D000228:Adenine Phosphoribosyltransferase / Q000172:deficiency / Q000235:genetics*; D000818:Animals; D002874:Chromosome Mapping; D003433:Crosses, Genetic; D004431:Ear, External; D005091:Exons; D005260:Female; D005347:Fibroblasts / Q000201:enzymology; D005808:Genes, Recessive; D014644:Genetic Variation; D006579:Heterozygote; D006801:Humans; D019656:Loss of Heterozygosity; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008815:Mice, Inbred Strains; D008817:Mice, Mutant Strains; D016296:Mutagenesis; D017354:Point Mutation; D011995:Recombination, Genetic; D012867:Skin / Q000166:cytology / Q000201:enzymology; D013601:T-Lymphocytes / Q000201:enzymology
D000228:Adenine Phosphoribosyltransferase