The in vivo Pig-a gene mutation assay, a potential tool for regulatory safety assessment.
The Pig-a (phosphatidylinositol glycan, Class A) gene codes for a catalytic subunit of the N-acetylglucosamine transferase complex involved in an early step of glycosylphosphatidyl inositol (GPI) cell surface anchor synthesis. Pig-a is the only gene involved in GPI anchor synthesis that is on the X-chromosome, and research into the origins of an acquired genetic disease involving GPI anchor deficiency (paroxysmal nocturnal hemoglobinuria) indicates that cells lacking GPI anchors, or GPI-anchored cell surface proteins, almost always have mutations in the Pig-a gene. These properties of the Pig-a gene and the GPI anchor system have been exploited in a series of assays for measuring in vivo gene mutation in blood cells from humans, rats, mice, and monkeys. In rats, flow cytometric measurement of Pig-a mutation in red blood cells requires microliter volumes of blood and data can be generated in hours. Spontaneous mutant frequencies are relatively low (<5 × 10(-6)) and rats treated with multiple doses of the potent mutagen, N-ethyl-N-nitrosourea, display Pig-a mutant frequencies that are close to the sum of the frequencies produced by the individual exposures. A general observation is that induced mutant frequencies are manifested earlier in reticulocytes (about 2 weeks after treatment) than in total red blood cells (about 2 months after exposure). Based on data from a limited number of test agents, the assay shows promise for regulatory applications, including integration of gene mutation measurement into repeat-dose toxicology studies.
Dobrovolsky
Vasily N
VN
U.S. Food and Drug Administration, National Center for Toxicological Research, Jefferson, Arkansas 72279, USA. vasily.dobrovolsky@fda.hhs.gov
Miura
Daishiro
D
Heflich
Robert H
RH
Dertinger
Stephen D
SD
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D005434:Flow Cytometry; D017261:Glycosylphosphatidylinositols / Q000235:genetics*; D006801:Humans; D051379:Mice; D009152:Mutagenicity Tests / Q000379:methods / Q000592:standards; D009154:Mutation; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism; D051381:Rats; D018570:Risk Assessment / Q000331:legislation & jurisprudence
D017261:Glycosylphosphatidylinositols; D017351:N-Acetylglucosaminyltransferases