Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Universidade de São Paulo, Ribeirão Preto, SP, Brasil.
Braz J Med Biol Res. 2010 Dec;43(12):1143-52. doi: 10.1590/s0100-879x2010007500126. Epub 2010 Nov 12.
5-Bromo-2'-deoxyuridine (BrdUrd) has long been known to interfere with cell differentiation. We found that treatment of Bradysia hygida larvae with BrdUrd during DNA puff anlage formation in the polytene chromosomes of the salivary gland S1 region noticeably affects anlage morphology. However, it does not affect subsequent metamorphosis to the adult stage. The chromatin of the chromosomal sites that would normally form DNA puffs remains very compact and DNA puff expansion does not occur with administration of 4 to 8 mM BrdUrd. Injection of BrdUrd at different ages provoked a gradient of compaction of the DNA puff chromatin, leading to the formation of very small to almost normal puffs. By immunodetection, we show that the analogue is preferentially incorporated into the DNA puff anlages. When BrdUrd is injected in a mixture with thymidine, it is not incorporated into the DNA, and normal DNA puffs form. Therefore, incorporation of this analogue into the amplified DNA seems to be the cause of this extreme compaction. Autoradiographic experiments and silver grains counting showed that this treatment decreases the efficiency of RNA synthesis at DNA puff anlages.
5-溴-2'-脱氧尿苷(BrdUrd)长期以来一直被认为会干扰细胞分化。我们发现,在多线染色体唾液腺 S1 区的 DNA 泡原基形成过程中,用 BrdUrd 处理黄粉虫幼虫,明显影响原基形态。然而,它不会影响随后向成虫阶段的变态。用 4 到 8mM BrdUrd 处理后,通常形成 DNA 泡的染色体位点的染色质仍然非常紧凑,DNA 泡扩张不会发生。在不同年龄注射 BrdUrd 会引发 DNA 泡染色质的压缩梯度,导致形成非常小的甚至几乎正常的泡。通过免疫检测,我们表明该类似物优先掺入 DNA 泡原基。当 BrdUrd 与胸苷混合注射时,它不会掺入 DNA,并且会形成正常的 DNA 泡。因此,这种类似物掺入扩增的 DNA 似乎是这种极端压缩的原因。放射自显影实验和银粒计数表明,这种处理会降低 DNA 泡原基处的 RNA 合成效率。