The micronucleus test and erythropoiesis. Effects of erythropoietin and a mutagen on the ratio of polychromatic to normochromatic erythrocytes (P/N ratio)

It is considered that a decrease of the ratio of polychromatic erythrocytes (PCE) to normochromatic erythrocytes (NCE) (P/N) in the micronucleus test is an indicator of bone marrow toxicity induced by mutagens. However, the exact meaning of fluctuation in the P/N ratio is not yet known. We have studied this point by counting the total number of erythrocytes and nucleated cells in the bone marrow following various treatments. The P/N ratio decreased gradually with time after administration of mitomycin C. Our data suggest that the decrease of P/N ratio was attributable to an increase in the numbers of the denominator, i.e. NCE, caused by rapid differentiation and multiplication or denucleation of erythroblasts which remained in the bone marrow instead of entering the peripheral blood stream. A decrease of P/N ratio was also observed in the early phase after administration of erythropoietin, an agent which induces differentiation and multiplication of erythroblasts. This phenomenon might result from an increase of PCE delivery into the blood circulation. However, following the initial decrease, the P/N ratio increased gradually 48 h after administration of erythropoietin. It is supposed that this increase probably resulted from an increase in PCE in the bone marrow due to the direct effects of erythropoietin on erythropoiesis. The drastic change in erythropoiesis in the bone marrow induced by either mutagen or erythropoietin treatment will affect the fluctuations of the P/N ratio or the number of micronucleated erythrocytes per non-micronucleated erythocytes in the micronucleus test. This contrasts with the original explanation for such fluctuations which attributed them to replenishment of the marrow by peripheral blood.