高K 对果蝇幼虫神经肌肉接头突触囊泡释放的超微结构研究

论文总字数：21752字

摘 要

突触囊泡内含有大量神经递质，具有突触传递的功能，是神经元突触前终末功能和结构的重要组成部分。突触囊泡功能受到影响会使得突触的传递功能发生障碍，从而导致各种精神类疾病和生理紊乱。研究表明，施加不同的刺激，突触囊泡释放的能力和运动方式也会随之改变。果蝇是一种重要的模式动物，但是目前关于果蝇外周神经肌肉接头突触囊泡在不同刺激下释放和回收的机制尚未被很好地阐明。本文选用模式动物果蝇，给予幼虫外周神经肌肉接头不同的处理条件，通过对处理后的结果进行观察分析，探索了果蝇外周突触囊泡在刺激条件下的结构特点和发育模式，从而为外周神经系统信号传递等分子机制提供形态学基础。同时将两种文献上提及的固定剂的效果进行了比较，选择了更为合适的电镜样品固定剂。

目的：观察果蝇突触扣结的超微结构，探索果蝇外周肌肉接头突触囊泡在K 和高温刺激下的结构、类型、数量变化。

方法：利用高温、高K 等条件对解剖出的w118野生型果蝇幼虫外周神经肌肉接头进行处理，然后通过常规电镜包埋技术对样品进行包埋，将包埋后的样品制成大量超薄切片，处理后在透射电镜下观察，分析所得图像。

结果：高温和高K 均能导致果蝇幼虫外周神经肌肉接头突触囊泡循环障碍，二者协同则导致循环发生严重障碍。突触囊泡的结构、类型、数量在刺激下会发生一系列变化，这些变化或许能够为研究突触传递提供分子学的基础。

关键词：果蝇,神经肌肉接头,超薄切片,突触囊泡

Ultrastructural study on synaptic vesicle release

from neuromuscular junction of Drosophila

larvae under high K condition

No：41113116 Student:Yizhen Chen

Supervisor:Guangming Gan

Abstract

Synaptic vesicles contain a large number of neurotransmitters, with synaptic transmission function, neuronal synaptic terminal function and an important part of the structure. Synaptic vesicle function is affected by the synaptic transmission function of the obstacles, resulting in a variety of mental illness and physical disorders. Studies have shown that the ability to exert a different stimulus, synaptic vesicle release, and movement patterns will change. Drosophila is an important model animal, but the mechanism of release and recovery of synaptic vesicles in the peripheral nerve of the Drosophila melanogaster is not well elucidated. In this paper, we selected the fruit flies of the model animals to give different treatment conditions of the larvae peripheral nerve muscular joints. Through the observation and analysis of the processed fruits, the structural characteristics and developmental patterns of the peripheral synaptosomes of Drosophila were studied. Peripheral nervous system signaling and other molecular mechanisms to provide morphological basis. At the same time, the fixation effect of two fixative mentioned in the literature was compared, and a more suitable electron microscope sample fixative was selected.

Objective:To observe the ultrastructure of synaptic buckling of Drosophila melanogaster to explore the structure, type and quantity of synaptic vesicles in the peripheral muscle of Drosophila melanogaster under K and high temperature stimulation.

Methods:The peripheral nerve muscular joints of w118 wild-type Drosophila larvae were treated by high temperature and high K conditions. Then, the samples were embedded by conventional electron microscopy embedding technique, and the coated samples were made into a large number of ultra-thin slices. To observe, collect the transmission electron microscope image, analysis and processing of the resulting image.

Results:Both high temperature and high K could lead to the synaptic vesicle circulation disorder in the peripheral nerve muscular joints of Drosophila larvae, and the synergistic effect could also lead to a serious circulatory disturbance. The structure, type and number of synaptic vesicles undergo a series of changes under stimulation, which may provide a molecular basis for studying synaptic transmission.

Key words:Drosophila, neuromuscular junction, ultra thin slices, synaptic vesicles

目 录

摘要..............................................................................................................................Ⅰ

Abstract........................................................................................................................Ⅱ

1 绪论

1.1突触的结构 1

1.2果蝇神经元K 通道 1

1.2.1果蝇K 通道的结构 1

1.2.2 K 通道在维持果蝇神经元兴奋性的作用 1

1.3突触囊泡循环与囊泡池 2

1.3.1即刻释放囊泡池 2

1.3.2循环池 2

1.3.3储存池 2

1.4果蝇幼虫外周神经肌肉接头突触扣结 3

1.4.1 Type I型 3

1.4.2 Type II型 4

1.4.3 Type III型 4

1.4.4其他分型方法 5

1.4.5突触扣结发育异常 6

1.5透射电镜 6

2 材料与方法

2.1实验材料 8

2.1.1果蝇品系 8

2.1.2培养基制备 8

2.1.3试剂的配制 8

2.2实验方法 8

2.3图像采集与整理 9

3 结果与讨论

3.1果蝇幼虫神经肌肉接头Type I型突触扣结的结构 10

3.2钾离子和高温对突触囊泡循环的影响 10

3.2.1高K 促进突触结构和突触囊泡循环异常 10

3.2.2高温促进突触结构和突触囊泡循环异常 11

3.2.3高温和高K 协同促进突触结构和突触囊泡循环异常 12

3.3固定剂选择的优化 13

3.4结论 13

3.5果蝇外周神经系统研究展望 13

参考文献 15

致谢 17

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