›› 2018, Vol. 61 ›› Issue (6): 733-740.doi: 10.16380/j.kcxb.2018.06.012

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基于改良石蜡切片技术的飞蝗胚胎浆膜表皮发育模式及形态变化观察

柳伟伟1,2, 李任建1,2, 付穗业1,2, 董卿1,2, 张敏1,  张学尧1, 张建珍1, 张婷婷1,*   

  1. (1. 山西大学应用生物学研究所, 农业有害生物综合治理山西省重点实验室, 太原 030006; 2. 山西大学生命科学学院, 太原 030006)
  • 出版日期:2018-06-20 发布日期:2018-06-20

Observation of the developmental pattern and morphological change of serosal cuticle in Locusta migratoria (Orthoptera: Acrididae) with an optimized paraffin section method

LIU Wei-Wei1,2, LI Ren-Jian1,2, FU Sui-Ye1,2, DONG Qing1,2, ZHANG Min1, ZHANG Xue-Yao1, ZHANG Jian-Zhen1, ZHANG Ting-Ting1,*    

  1. (1. Shanxi Key Laboratory of Integrated Pest Management in Agriculture, Institute of Applied Biology, Shanxi University, Taiyuan 030006, China; 2. College of Life Science, Shanxi University, Taiyuan 030006, China)
  • Online:2018-06-20 Published:2018-06-20

摘要: 【目的】通过优化飞蝗Locusta migratoria胚胎石蜡切片制备技术,研究飞蝗胚胎发育时期浆膜表皮变化规律。【方法】通过优化飞蝗胚胎固定前处理、洗涤、脱水和透明等步骤,改良胚胎期石蜡切片制备技术;制备飞蝗胚胎发育第3-10天的石蜡切片,通过HE染色观察浆膜表皮发育规律;通过制备飞蝗发育第6-8天从卵孔端至卵尾端的纵切切片,观察飞蝗胚胎转旋时期的形态变化。【结果】飞蝗胚胎期石蜡切片技术优化后的操作步骤为:固定前NaClO预处理和打孔、洗涤30 min、脱水30 min和透明30 min。通过此方法可获得表皮结构完整且清晰的胚胎期组织切片。在30℃条件下,飞蝗胚胎浆膜表皮及几丁质沉积形成于胚胎发育的第5天,在第8天时开始降解。飞蝗胚胎的转旋发生在第6-7天,伴随着浆膜和浆膜表皮的形态变化。【结论】本研究优化了飞蝗胚胎石蜡切片制备步骤,揭示了飞蝗胚胎浆膜表皮及胚胎转旋的发育模式,为昆虫胚胎发育的研究提供了基础资料。

关键词: 飞蝗, 石蜡切片改良, HE染色, 胚胎, 浆膜表皮, 胚胎转旋

Abstract: 【Aim】 The aim of this study is to optimize the paraffin section technology and to examine the developmental pattern of serosal cuticle in the early embryonic stage of Locusta migratoria. 【Methods】 The paraffin section technology for L. migratoria embryo was modified on the steps of fixation, washing, dehydration and clearing. The developmental process of serosal cuticle was analyzed with hematoxylin-eosin staining (HE staining) and fluorescent staining following paraffin section of day 3-10 embryos of L. migratoria. The katatrepsis of embryo from micropyle to tail was observed by HE staining of the paraffin sections of L. migratoria from day 6-8 embryos, which were cut longitudinally. 【Results】 The optimized paraffin section steps for L. migratoria embryo included: pretreating the egg with NaClO and punching it before fixation, then washing, dehydrating and clearing the fixed egg for 30 min, respectively. Integrative and clear sections of the whole epidermis structure of the embryonic stage were obtained by using our optimized paraffin section technology. The serosal cuticle formation and chitin deposition were completed on day 5 and began to degrade on day 8 during the embryonic stages at 30℃. The katatrepsis occurred from day 6 to day 7 in the embryonic stages, accompanied by the morphological change of serosa and serosal cuticle. 【Conclusion】 We optimized the paraffin section technology for L. migratoria embryo, and revealed the developmental pattern of serosal cuticle and katatrepsis. This study lays a foundation for studying the metabolisms of embryonic development in L. migratoria.

Key words: Locusta migratoria, paraffin section modification, HE staining, embryo, serosal cuticle, katatrepsis