Optogenetics in the teaching laboratory: using channelrhodopsin-2 to study the neural basis of behavior and synaptic physiology in Drosophila
Optogenetics in the teaching laboratory: using channelrhodopsin-2 to study the neural basis of behavior and synaptic physiology in Drosophila
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DOI：
10.1152/advan.00125.2010
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发表时间：
2011-03-01
影响因子：
2.1
通讯作者：
Johnson, Bruce R.
中科院分区：
文献类型：
--
作者：
Pulver, Stefan R.;Hornstein, Nicholas J.;Johnson, Bruce R.
Pulver SR、Hornstein NJ、Land BL、Johnson BR。教学实验室中的光遗传学：利用视紫红质通道蛋白2研究果蝇行为和突触生理学的神经基础。高级生理教育 35：82-91，2011； doi:10.1152/advan.00125.2010.-在这里，我们将果蝇神经遗传学和“光遗传学”的最新进展纳入神经科学实验室练习中。我们使用光激活离子通道视紫红质-2 (ChR2) 和组织特异性基因表达技术来研究果蝇幼虫行为的神经基础。我们使用廉价、易于使用的系统设计并实施了练习，以提供具有精细时间控制的蓝光脉冲。学生首先检查激活果蝇幼虫谷氨酸能神经元的行为影响，然后记录幼虫神经肌肉接头 (NMJ) 处 ChR2 激活介导的兴奋性连接电位 (EJP)。电诱发 EJP 和光诱发 EJP 的比较表明，光诱发 EJP 的幅度和时间进程与电神经刺激产生的幅度和时间进程没有显着差异。这些练习向学生介绍了用于远程操纵神经活动的新遗传技术，并简化了在果蝇幼虫 NMJ 上记录 EJP 的过程。在果蝇中使用 ChR2 进行的研究工作相对较少，因此学生有机会测试新颖的假设并为科学记录做出切实的贡献。对学生体验的定性和定量评估表明，这些练习有助于传达突触传递原理，同时还促进遗传学、细胞生理学和动物行为的综合和探究性研究。
Pulver SR, Hornstein NJ, Land BL, Johnson BR. Optogenetics in the teaching laboratory: using channelrhodopsin-2 to study the neural basis of behavior and synaptic physiology in Drosophila. Adv Physiol Educ 35: 82-91, 2011; doi:10.1152/advan.00125.2010.-Here we incorporate recent advances in Drosophila neurogenetics and "optogenetics" into neuroscience laboratory exercises. We used the light-activated ion channel channelrhodopsin-2 (ChR2) and tissue-specific genetic expression techniques to study the neural basis of behavior in Drosophila larvae. We designed and implemented exercises using inexpensive, easy-to-use systems for delivering blue light pulses with fine temporal control. Students first examined the behavioral effects of activating glutamatergic neurons in Drosophila larvae and then recorded excitatory junctional potentials (EJPs) mediated by ChR2 activation at the larval neuromuscular junction (NMJ). Comparison of electrically and light-evoked EJPs demonstrates that the amplitudes and time courses of light-evoked EJPs are not significantly different from those generated by electrical nerve stimulation. These exercises introduce students to new genetic technology for remotely manipulating neural activity, and they simplify the process of recording EJPs at the Drosophila larval NMJ. Relatively little research work has been done using ChR2 in Drosophila, so students have opportunities to test novel hypotheses and make tangible contributions to the scientific record. Qualitative and quantitative assessment of student experiences suggest that these exercises help convey principles of synaptic transmission while also promoting integrative and inquiry-based studies of genetics, cellular physiology, and animal behavior.