In vivo rodent micronucleus assay: protocol, conduct and data interpretation.

In vivo rodent micronucleus assay has been widely used to detect genotoxicity. Evaluation of micronucleus induction is the primary in vivo test in a battery of genotoxicity tests and is recommended by the regulatory agencies around the globe to be conducted as part of product safety assessment. The assay, when performed appropriately, detects both clastogenicity and aneugenicity. Methods for performing micronucleus evaluation have evolved since its initial description in the 1970s. In recent years, the focus has been directed toward improving micronucleus detection with high efficiency by proposing data-based recommendations to the standard initial protocol design. Such improvements include, e.g., the use of appropriate harvest time(s), inclusion of one or both sexes, number of doses tested, limit dose, integrating micronucleus assessment into the routine toxicology studies, use of fluorescent staining, automation of micronucleus detection and assessment of micronuclei in multiple tissues. This protocol paper describes: the mechanism of micronucleus formation, a generalized protocol for manual detection, enumeration of micronuclei, and data interpretation in light of published information thus far, on the regulatory aspects of this assay. Certain recent protocol issues that are practical in nature are equally valid in relation to standard manual method and provide robust database, which are also included for consideration. It is expected that such improvements of the protocol will continue to drive the utility of this assay in the product safety assessment.