title: Report on the rat Pig-a assay using an anti-rat erythroid marker HIS49 antibody in a single dose study of 1,2-dimethylhydrazine.
abstract: As part of a collaborative study in the Mammalian Mutagenicity Study (MMS) Group of the Japanese Environmental Mutagen Society (JEMS), we investigated the in vivo genotoxicity profile of 1,2-dimethylhydrazine (DMH) using a Pig-a assay in total red blood cells (RBC Pig-a assay) or a reticulocyte Pig-a assay (PIGRET assay). We also assessed the genotoxic potential of DMH using both a bone marrow micronucleus test and a liver comet assay as follow-up studies. Single administration of 25, 50, 100mg/kg DMH to male rats did not show time- or dose-related increases in Pig-a mutant frequency (MF) in either the RBC Pig-a or PIGRET assays up to 4 weeks after treatment. The bone marrow micronucleus test under the same dose levels was judged positive, while the liver comet assay was judged inconclusive due to the high number of hedgehogs. Re-evaluation of the rat liver comet assay at lower dose levels (4, 10, and 25mg/kg DMH) showed a dose-related increase in%DNA in tail. Taken together, DMH showed a positive response in both the bone marrow micronucleus test and liver comet assay, while the increases in Pig-a MF in both the RBC Pig-a and PIGRET assays could hardly be detected after single dosing. These results suggest that DMH provides different genotoxicity outcomes depending on the endpoint following acute in vivo dosing.
authors.affiliation: Pharmaceutical Development Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Limited, 4-3-2 Asahigaoka, Hino-shi, Tokyo, 191-8512, Japan.
authors.forename: Satsuki
authors.initials: S
authors.lastname: Chikura
authors.affiliation: Pharmaceutical Development Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Limited, 4-3-2 Asahigaoka, Hino-shi, Tokyo, 191-8512, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Okada
authors.affiliation: Pharmaceutical Development Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Limited, 4-3-2 Asahigaoka, Hino-shi, Tokyo, 191-8512, Japan. Electronic address: t.kimoto@teijin.co.jp.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Kimoto
authors.affiliation: Pharmaceutical Development Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Limited, 4-3-2 Asahigaoka, Hino-shi, Tokyo, 191-8512, Japan.
authors.forename: Hideshi
authors.initials: H
authors.lastname: Kaneko
authors.affiliation: Pharmaceutical Development Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Limited, 4-3-2 Asahigaoka, Hino-shi, Tokyo, 191-8512, Japan.
authors.forename: Daishiro
authors.initials: D
authors.lastname: Miura
authors.affiliation: Pharmaceutical Development Research Laboratories, Teijin Institute for Bio-medical Research, Teijin Pharma Limited, 4-3-2 Asahigaoka, Hino-shi, Tokyo, 191-8512, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kasahara
mesh_terms: D019813:1,2-Dimethylhydrazine / Q000633:toxicity*; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics*; D009152:Mutagenicity Tests / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D008565:Membrane Proteins; C080086:phosphatidylinositol glycan-class A protein; D019813:1,2-Dimethylhydrazine
keywords: Comet assay; Flow cytometry; Micronucleus test; Pig-a assay