BrdU抗体染色技术检测昆虫器官的细胞增殖

Revealing the proliferation rate in insect organs by BrdU antibody staining

BrdU(5-bromo-2'-deoxyuridine)是一种人工合成的核苷酸类似物,常用于标记活体组织的增殖细胞。它的结构类似于胸腺嘧啶,在细胞分裂的S期,可以取代胸腺嘧啶而插入正在复制的细胞DNA中。通过免疫组化手段,用BrdU抗体检测整合在细胞DNA中的BrdU分子,可以反映出细胞周期的活力,即细胞增殖的速率。本文介绍一种优化的BrdU染色流程,用来标记昆虫小器官的细胞增殖速率。通过这种技术,我们重新评估了Dpp信号通路中的转录抑制因子Brinker在翅芽发育过程中调控细胞增殖的作用,发现它并不是以前所认为的在翅囊区是一种生长抑制因子。

A synthetic nucleoside, Bromodeoxyuridine （5bromo2′deoxyuridine, BrdU）, is commonly used to detect proliferating cells in living tissues. This compound is an analogue of thymidine and is incorporated into newly synthesized DNA of S phase cells by being substituted for thymidine during DNA replication. Immunohistochemical staining of BrdU is then used to detect the incorporated BrdU and thereby reveal the actively replicating cells. This article introduces an optimized BrdU protocol for detecting cell proliferation in small insect organs. Using this technique, we reassessed the role of a transcriptional repressor, Brinker, in the Dpp signaling pathway controlling growth regulation during wing disc development. We found that, contrary to previous belief, Brinker is not a growth repressor in the wing pouch.