It is generally believed that, during Xenopus laevis oogenesis, polymerase I transcription is high in the early vitellogenic oocytes (stages III and IV) and very low in later stages. We used a combination of RNA labeling, nuclease S1 protection assays, Northern blot, and half-life measurement of preribosomal RNA to reinvestigate the pattern of polymerase I activity during oogenesis. Unexpectedly, when we compared the amount of 40S pre-rRNA produced in stages IV and VI by direct labeling or with a probe that hybridizes with the 5' external transcribed spacer, we found a high level of 40S pre-rRNA in stage VI oocytes. This precursor ribosomal RNA transcribed in stage VI oocytes is processed to give the matured 18S and 28S species. These results suggest that the activity of RNA polymerase I in stage VI oocytes is similar or very close to that found in stage IV, which is probably required to maintain the huge number of ribosomes during oogenesis.