Gu Shi-Hong, Chow Yien-Shing
Department of Zoology, National Museum of Natural Science, 1 Kuan Chien Road, Taichung, Taiwan 404-19, R. O. C.
J Exp Zool A Comp Exp Biol. 2005 Apr 1;303(4):249-58. doi: 10.1002/jez.a.142.
DNA synthesis in prothoracic gland cells of the silkworm, Bombyx mori, was studied immunocytochemically after in vivo labeling with 5-bromo-2'-deoxyuridine (BrdU), and its developmental changes during the 3rd, 4th, and last larval instars were examined. During the early stages of both the 3rd and 4th larval instars, a dramatic increase in the number of DNA-synthesizing cells of the prothoracic glands was detected. However, during the latter stages of each instar, the number of DNA-synthesizing cells greatly decreased. The determination of glandular protein content showed that dramatic increases occurred during the latter stages of each larval instar. Comparison of changes in prothoracic gland cell DNA synthesis with ecdysteroidogenic activity showed that the increase in DNA synthesis precedes ecdysteroidogenesis. The cellular mechanism underlying changes in prothoracic gland cell DNA synthesis during the last two larval instars was further analyzed by determining the in vitro DNA synthesis of the glands, their responsiveness to hemolymph growth factors, and changes in the growth-promoting activity of hemolymph during development. It was found that both growth factors and the responsiveness of the prothoracic gland cells to growth factors from hemolymph may play roles in regulating DNA synthesis of gland cells.
在用5-溴-2'-脱氧尿苷(BrdU)进行体内标记后,通过免疫细胞化学方法研究了家蚕前胸腺细胞中的DNA合成,并检测了其在幼虫第三龄、第四龄和最后一龄期间的发育变化。在第三龄和第四龄幼虫的早期阶段,检测到前胸腺中DNA合成细胞的数量急剧增加。然而,在每一龄的后期,DNA合成细胞的数量大幅减少。腺体蛋白质含量的测定表明,在每一龄幼虫的后期都出现了急剧增加。将前胸腺细胞DNA合成的变化与蜕皮甾体生成活性进行比较,发现DNA合成的增加先于蜕皮甾体生成。通过测定腺体的体外DNA合成、它们对血淋巴生长因子的反应以及发育过程中血淋巴促生长活性的变化,进一步分析了最后两龄幼虫期间前胸腺细胞DNA合成变化的细胞机制。结果发现,生长因子以及前胸腺细胞对血淋巴中生长因子的反应性可能在调节腺体细胞的DNA合成中发挥作用。