The Pig-a gene mutation assay, a powerful tool for evaluating in vivo genotoxicity, is based on flow cytometric enumeration of red blood cells (RBCs), which are deficient in glycosylphosphatidylinositol anchored proteins caused by mutation(s) in the Pig-a gene. Various approaches for measuring cells with mutated Pig-a gene have been developed. The Pig-a assay targeting concentrated reticulocytes - the PIGRET assay - has the potential to detect genotoxicity in early stages of the study. To verify the potential and usefulness of the PIGRET assay for short-term testing, we conducted a joint research with the Mammalian Mutagenicity Study (MMS) Group of the Japanese Environmental Mutagen Society. As part of this study, we evaluated the genotoxicity of a single oral administration of acrylamide (AA) at 25, 50, 100, 137.5, and 175mg/kg using the PIGRET and Pig-a assays targeting RBCs (RBC Pig-a assay) at 7, 14, and 28 days after dosing. Toxic effects induced by AA, such as hind limb weak-paralysis, reduction of body weight gain, and reticulocytosis, were observed in AA-treated groups. However, we detected no significant increases in Pig-a mutant frequencies using either the PIGRET or RBC Pig-a assay. Therefore, we concluded that the genotoxicity of AA could not be detected by these assays under our experimental conditions.

中文翻译：

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在大鼠中使用 RBC Pig-a 和 PIGRET 试验通过单次口服剂量评估丙烯酰胺的致突变性。

Pig-a 基因突变检测是一种评估体内遗传毒性的强大工具，它基于红细胞 (RBC) 的流式细胞计数，红细胞缺乏由 Pig-a 基因突变引起的糖基磷脂酰肌醇锚定蛋白. 已经开发了多种测量具有突变 Pig-a 基因的细胞的方法。以浓缩网织红细胞为目标的 Pig-a 试验——PIGRET 试验——有可能在研究的早期阶段检测基因毒性。为了验证 PIGRET 检测在短期测试中的潜力和实用性，我们与日本环境诱变剂协会的哺乳动物致突变性研究 (MMS) 小组进行了联合研究。作为本研究的一部分，我们评估了单次口服丙烯酰胺 (AA) 的遗传毒性，浓度分别为 25、50、100、137.5、在给药后 7、14 和 28 天，使用针对 RBC 的 PIGRET 和 Pig-a 测定（RBC Pig-a 测定）和 175mg/kg。在AA治疗组中观察到AA引起的毒性作用，例如后肢无力麻痹、体重增加减少和网状细胞增多症。然而，我们使用 PIGRET 或 RBC Pig-a 检测没有检测到 Pig-a 突变频率的显着增加。因此，我们得出结论，在我们的实验条件下，这些测定无法检测到 AA 的遗传毒性。我们使用 PIGRET 或 RBC Pig-a 检测未检测到 Pig-a 突变频率的显着增加。因此，我们得出结论，在我们的实验条件下，这些测定无法检测到 AA 的遗传毒性。我们使用 PIGRET 或 RBC Pig-a 检测未检测到 Pig-a 突变频率的显着增加。因此，我们得出结论，在我们的实验条件下，这些测定无法检测到 AA 的遗传毒性。