蜜蜂全脑解剖、腺体分离及不同日龄组织形态比较

Whole Brain Anatomy,Glandular Separation and Histological Comparison of Honeybee at Different Age

蜜蜂作为一种重要的社会性昆虫,具有复杂的学习记忆和精确的信息交流能力。蜜蜂大脑是各种信息的处理中心,是模式生物神经生物学的重要研究对象。精确解剖蜜蜂脑是研究神经生物学的关键,但由于蜜蜂脑体积小且被几丁质外壳包裹,完整解刨脑部并进行脑内分区一直是技术难题。为解决蜜蜂脑解剖难度大、速度慢和质量差的问题,我们探索和改进蜜蜂脑解剖方法,实现了快速高效的解剖蜜蜂脑及其亚分区,为深入研究蜜蜂脑神经生物学提供帮助。取新鲜或CO2麻醉的蜜蜂,用2根昆虫针穿过胸部正面固定于蜡盘,在显微镜下用尖头镊子划开复眼后方的几丁质外壳,掀开脑壳,取出全脑放在含有蛋白酶/RNA酶抑制剂溶液的载玻片上,然后进行组织分区。经过练习掌握方法后,可以快速准确解剖蜜蜂脑,并获得蘑菇体、咽下腺、上颚腺、视神经叶和嗅神经叶等亚器官结构,从解剖到分区可以在2分钟内完成,且样品的外部形态完整,质量好。蜜蜂头部的器官解剖结果显示,出房蜂、哺育蜂和采集蜂3个日龄蜜蜂脑、咽下腺、上颚腺、唾液腺均有明显差异,这与其器官发育和劳工分配具有密切关系。

As an important kind of social insect,honeybee has complex abilities of learning,memory,and communication of information.The brain of honeybee is the processing center of various information.It is gradually becoming the model organism in the research field of neurobiology because of the specific strengths of honeybee’s brain.Brain anatomy is the first step in studying the brain of honeybee.However,the small volume and the chitinous cover of honeybee’s brain make technological diffi culties to complete the anatomy of the brain as well as the partition of inner brain.To get over the diffi culties including the slow speed and the poor quality of the brain tissue,we perform a complete technology to dissect the bee brain and the inner brain partition and provide help to the follow-up in-depth research of honeybee’s neuroscience by exploring and improving the methods of brain anatomy.Firstly,the fresh honeybee or honeybee that was smoked by CO2 was frontally fi xed to the wax tray by using two insect needles to stick through the breast.Secondly,the chitin shell was gashed from the behind of the compound eye and the skull was opened by tweezers.Then,the whole brain was taken out and put on the glass slide with PBS solution contained protease/RNA polymerase inhibitor.After mastering the methods by practicing,the process from anatomy to partition could be fi nished and the sub organ including the mushroom body,hypopharyngeal gland,mandibular gland,optic lobe,and olfactory lobe could be gotten in two minutes.More importantly,the external form of the brain sample was completely preserved with adequate quality.The brain of honeybee is the center of controlling behaviors and physiological activities.The images of organ samples from the newly emerged bee,nurse bee,and foraging bee have shown signifi cant differences,which are closely related to their organ development and labor allocation.