›› 2017, Vol. 60 ›› Issue (12): 1361-1366.doi: 10.16380/j.kcxb.2017.12.001

• 研究论文 •    下一篇

利用G-TRACE技术追踪果蝇后肠肠细胞谱系的发育模式

张徐波1,*, 张月1,2, 董玮1, 李开霞1, 吴海花1, 张敏1   

  1. (1. 山西大学应用生物学研究所, 太原 030006; 2. 山西大学生命科学学院, 太原 030006)
  • 出版日期:2017-12-20 发布日期:2017-12-20

Lineage tracing of Drosophila hindgut enterocytes by G-TRACE

ZHANG Xu-Bo1,*, ZHANG Yue1,2, DONG Wei1, LI Kai-Xia1, WU Hai-Hua1, ZHANG Min1   

  1. (1. Institute of Applied Biology, Shanxi University, Taiyuan 030006, China; 2. College of Life Science and Technology, Shanxi University, Taiyuan 030006, China)
  • Online:2017-12-20 Published:2017-12-20

摘要: 【目的】果蝇是完全变态昆虫,蛹期经历了幼虫组织解离和成虫组织重塑的过程。本研究旨在利用细胞谱系追踪方法G-TRACE(Gal4 technique for real-time and clonal expression)这一新的遗传学技术,检测果蝇幼虫后肠肠细胞在蛹期发育过程中是否发生细胞迁移。【方法】采用黑腹果蝇Drosophila melanogaster engrailed-Gal4(en-Gal4)品系和G-TRACE品系杂交,并引入tub-gal80ts控制Gal4的开启时间,分别在果蝇幼虫期和蛹期进行细胞谱系追踪。幼虫期追踪:亲代产卵后将卵置于30℃培养,3龄中期转入18℃培养,成虫羽化1 d内进行检测。蛹期追踪:亲代产卵后将卵置于18℃培养,在蛹期不同发育阶段转入30℃培养,待虫体羽化后检测成虫肠道。【结果】当在果蝇幼虫期启动细胞谱系追踪,在蛹期停止追踪,发现中肠靠近中后肠边界处以及马氏管存在绿色肠细胞。而当在果蝇幼虫期关闭细胞谱系追踪,在蛹期开始追踪,则发现虫体中肠各部位及马氏管分布着绿色肠细胞。en基因在果蝇蛹期肠道中表达。【结论】结果表明,在果蝇蛹形成过程中,后肠的部分肠细胞迁移至中肠和马氏管,参与中肠和马氏管的重塑。本研究对于探索昆虫在变态发育过程中成虫器官的重塑机制具有重要的意义。  

关键词: 果蝇, 变态昆虫, 肠道, 细胞谱系追踪, 细胞迁移, G-TRACE

Abstract: 【Aim】 Drosophila is one of holometabolous insects. It undergoes the process of dissociation of old tissues and remodeling of adult tissues at the pupal stage. The objective of this study is to investigate whether hindgut enterocytes of Drosophila migrate into the midgut during metamorphosis by G-TRACE (Gal4 technique for real-time and clonal expression) which is a new genetic technique. 【Methods】 engrailed-Gal4 (en-Gal4) line and lineage-tracing line (G-TRACE) of Drosophila melanogaster were hybridized, and tub-gal80ts was introduced to temporally control Gal4 activity. The cell lineage was traced at the larval and pupal stages, respectively. For larval stage tracing, eggs were cultured at 30℃ after egg-laying by the parental generation, and the mid 3rd instar larvae were shifted to 18℃. The adult guts were detected within 1 d after eclosion. For pupal stage tracing, eggs were cultured at 18℃ after egg-laying by the parental generation, pupae were shifted to 30℃ at different stages, and adult guts were detected after eclosion. 【Results】 In larval stage tracing of Drosophila, green intestinal cells appeared in the posterior section of midgut, which is adjacent to the hindgut-midgut boundary and Malpighian tubules. In pupal stage tracing of Drosophila, green intestinal cells appeared at different sections in the midgut and Malpighian tubules, and engrailed gene was expressed in Drosophila intestine at the pupal stage. 【Conclusion】 These results suggest that during pupa formation, part of the hindgut cells migrate into the midgut or Malpighian tubules and are involved in the reformation of adult midgut or Malpighian tubules. This study is of important significance in understanding the mechanisms of insect organ remodeling during metamorphosis.

Key words: Drosophila, holometabolous insect, intestine, cell lineage tracing, cell migration, G-TRACE