ARTICLE

THE EFFECTS OF 5-BROMODEOXYURIDINE ON YOLK SAC ERYTHROPOIESIS IN THE CHICK EMBRYO

Yasusada Miura ¹ and Fred H. Wilt ¹

¹ From the Department of Zoology, University of California, Berkeley, California 94720.

Dr. Miura's permanent address is III Department of Internal Medicine, Faculty of Medicine, Tokyo University, Hongo, Bunkyo-ku, Tokyo

The effects of the thymidine analog, 5-bromodeoxyuridine (BUdR), on the formation of red cells in the yolk sac of the chick embryo were examined. The prospective area opaca vasculosa from a definitive primitive streak embryo was excised, disaggregated, and deposited into a cell clump, and the cell clump was placed in organ culture. Hemoglobin synthesis is detectable after about 16 hr in culture. The formation of erythropoietic foci and incorporation of ⁵⁵Fe into heme were used to measure the extent of erythropoiesis. Exposure to 40 µg/ml of BUdR within 6 hr after explantation almost completely eliminated red cell formation; subsequent transfer to thymidine medium showed that the inhibition was reversible, and there was no histological evidence of analog toxicity. Between 6 and 12 hr after initiation of organ culture, the tissue became completely refractory to BUdR. DNA synthesis, as monitored by thymidine-³H and BUdR-³H pulses, was extensive both during and after the period of BUdR sensitivity. Hence, during both BUdR sensitive and insensitive periods the analog was incorporated into DNA of cells which had not yet synthesized hemoglobin. It is proposed that between 6 and 12 hr a crucial regulatory event for terminal differentiation is perturbed by the presence of BUdR in the chromosomes.

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