在使用单一感器录音昆虫气味的嗅觉感官神经元的电生理反应,可测。在这个视频文章,我们将演示如何执行单一飞醋的触角感器录音(。<em>果蝇</em>)和疟疾的蚊子的上颌触须(<em>按蚊</em>)。
昆虫的嗅觉寻找食物,配偶,天敌和产卵场所3,。昆虫的嗅觉感觉神经元(OSNs)被称为感受器的感觉毛,其中包括嗅觉器官表面封闭。每个感器的表面覆盖着微小的孔隙,通过气味传递和化解感器淋巴液的液体,沐浴安置在一个给定的感器OSNs的感官树突。通过OSN树突表达气味受体(OR)的蛋白质,它在昆虫气味门控离子通道4,5的功能。互动的ORs要么增加气味或减少通过OSN基础射速。这在动作电位形成的神经元活动体现的质量,强度和时间特征的气味6,7的第一个代表性。
鉴于方便地访问这些感觉毛,它可以执行从单一OSNs引入感器淋巴结记录电极,参比电极放置在昆虫的眼睛或身体的淋巴结外的录音。在果蝇 ,一至四个OSNs感器的房子,但每个OSN通常会显示一个特征的尖峰幅度。穗排序技术使人们有可能分配扣球个别OSNs的反应。这种单一感器记录(SSR)技术监控潜在OSNs 1,2,8受体活性产生的尖峰电脉冲感器淋巴液和参比电极之间的区别。在尖峰数量变化的气味代表的气味编码在昆虫细胞的基础。这里,我们描述的制备方法,目前在我们的实验室用来执行对果蝇和按蚊的SSR,并显示在感器特定方式的气味引起的代表痕迹。
用于生物识别的食物来源,潜在的队友,和食肉动物的嗅觉线索。嗅觉感觉神经元(OSNs)之间的外部刺激,并进一步处理信息的大脑中心的第一个接力中心。在果蝇和按蚊 ,OSNs方便,可以监视他们的电活动而刺激的气味喷。
单感器记录(SSR)技术的解释,该视频已被广泛应用于从OSNs记录和研究它们的电反应,大量的气味6, 7。嗅觉受体(ORS)6,11…
Material Name | タイプ | Company | Catalogue Number | Comment |
---|---|---|---|---|
Paraffin oil | Odors | Fluka | 76235 | |
High purity odors (>98%) | Odors | Sigma-Aldrich | Methyl acetate #296996 1-octen-3-ol #74950 |
|
Filter paper strips | Odors | Fisherbrand | 05-714-1 | Chromatography paper |
Connectors | Odors | Cole-Parmer | EW-06365-40 | 1/16×1/8″ |
Glass vials | Odors | Agilent Technologies | 5182-0556 | |
Air line plastic tubing | Odor Delivery | Python Products | 500PAL | |
1 serological pipette | Odor Delivery | Corning | 4101 | 10 mL |
Plastic tubing | Odor Delivery | Cole-Parmer | EW-06418-0 | 0.050″x0.090″OD |
Disposable borosilicate glass Pasteur pipettes | Odor Delivery | FisherBrand | 13-678-20A | 5-3/4 inches |
Programmable stimulus controller | Odor Delivery | Syntech | CS-55 | |
Anti-vibration table | Electrophysiology Equipment | TMC | 63533 | 36”Wx30”Dx29”H |
Faraday cage | Electrophysiology Equipment | TMC | MI8133303 | |
Inverted microscope | Electrophysiology Equipment | Nikon | E600FN ECLIPSE | Recording microscope |
10x and 100x objectives | Electrophysiology Equipment | Nikon | 10x Plan Fluor 100x L Plan | |
Dissecting microscope | Electrophysiology Equipment | Nikon | EZ645 | electrode sharpening/insect prep microscope |
Magnetic stands | Electrophysiology Equipment | Newport | MODEL 150 | |
IDAC | Electrophysiology Equipment | Syntech | IDAC-4 | |
Acquisition software | Electrophysiology Equipment | Syntech | Autospike | |
1 macromanipulator | Electrophysiology Equipment | NARISHIGE | MN-151 | Joystick manipulator Used for positioning reference electrode |
1 micromanipulator | Electrophysiology Equipment | EXFO | PCS-6000 | Used for positioning recording electrode |
Crocodile clip | Electrophysiology Equipment | Pomona | AL-B-12-0 | |
Electric cable | Electrophysiology Equipment | Pomona | B-36-0 | Test Cable Assembly |
2 electrode holders | Electrophysiology Equipment | Syntech | N/A | Electrode holders (set of 2) for tungsten wire electrode |
AC probe | Electrophysiology Equipment | Syntech | N/A | Universal single ended probe (10xAC) |
Tungsten electrodes | Electrophysiology Equipment | Microprobes | M210 | straight tungsten rods, 0.005“x3“ |
Potassium hydroxide | Electrophysiology Equipment | Sigma-Aldrich | 221473 | |
Syringe | Electrophysiology Equipment | BD | 301625 | 20 mL |
Power supply | Electrophysiology Equipment | WILD HEERBRUGG 6V 40W | e.g MTR32 | |
Vertical puller | Insect prep | Narishige | PB-7 | |
Razor blade | Insect prep | VWR | 55411-050 | |
Dental wax | Insect prep | Patterson | 091-1503 | |
Microscope slide | Insect prep | FisherBrand | 12-550A | |
Cover glass | Insect prep | FisherBrand | 12-541A | 18X18 #1.5 |
Polypropylene mesh | Insect prep | Small Parts inc. | CMP-0500-B | |
Glass electrode | Insect prep | Frederick Haer & Co. | 27-32-0-075 | Capillary tubing borosilicate 1.5mm OD x 1.12mm ID x 75 mm |
Double-sided tape (3M) | Insect prep | 3M | MMM6652P3436 | Double-sided tape (3M) |
Forceps | Insect prep | Fine Science Tools | 021×0053 | Dumont #5 Mirror Finish Forceps |
Small plastic cup | Insect prep | VWR | 89009-662 | 7 x 5.7 (23/4 x 21/4) |
Electric aspirator | Insect prep | Gempler’s | RHM200 |