Department of Entomology, University of California, Davis
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Syed, Z., Leal, W. S. Electrophysiological Measurements from a Moth Olfactory System. J. Vis. Exp. (49), e2489, doi:10.3791/2489 (2011).
Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively. These approaches have been exploited for getting a better understanding of insect chemical communication. The identified stimuli can then be used as either attractants or repellents in management strategies for insect pests.
2. Electrophysiological Preparation
3. Chemical Stimuli and Stimulation Protocol
4. Representative Results
Antennae of restrained moths (Figure 1) stimulated with female pheromone component, (Z,Z)-11,13-hexadecadienal generate robust dose dependent responses (Figure 2). This major pheromone component also elicits dose dependent excitatory responses from long trichodea sensilla (Figure 3A) as displayed in traces of single unit recordings (Figure 3B).
Figure 1. Live navel orangeworm moth restrained and antenna exposed
Figure 2. Dose dependent EAG responses elicited in response to (Z,Z)-11,13-hexadecadienal, a major female pheromone constituent.
Figure 3. Sensilla trichoid in male navel orangeworm moth respond to (Z,Z)-11,13-hexadecadienal in a dose dependent manner. (A) Antenna of male navel orangeworm moth is multi-segmented and each segment is adorned with a large number of hair like structures, sensilla. A scanning electron micrograph, showing details in the inset (Scale bars are 200 and 50 μM, respectively). (B) Extracellular single-unit recordings from a trichoid sensillum.
This novel method developed for restraining a live navel orangeworm moth to measure olfactory signals has proved to be robust and very reliable. We routinely employ this method for isolating and identifying novel attractants from natural host substrates like almonds and pistachios.
No conflicts of interest declared.
Supported by USDA-NIFA/AFRI 2010-65105-20582, NSF 0918177, and Bedoukian Research Incorporation.
|Microscope||Olympus Corporation||BX51WI model|
|Stereo microscope||Leica Microsystems||MZ75|
|1.0 mm borosilicate capillary tubing with filament||World Precision Instruments, Inc.||1B100F-3|
|Micropipette puller||Sutter Instrument Co.||P-97|
|Stimulus controller||Syntech||CS-55 model|
|High Impedance pre-amplifiers (Universal Single ended probe)||Syntech|
|Amplifier / data-acquisition system (USB-IDAC)||Syntech|
|EAG Micromanipulator MP-12||Syntech|
|(Z,Z)-11,13-hexadecadienal||Bedoukian Research Inc, CT.|
|Whatman filter paper||Whatman, GE Healthcare||1001070|
|5 mL polypropylene syringe||BD Biosciences||309633|
|pipette tip (200 μL)||USA Scientific, Inc.||1111-0806|
|669 mL lunchbox,13 x 13 cm; height, 4.5 cm,||Rubbermaid|
|Thirsty Ultra Absorbent, 27.9 x 27.9 cm||Safeway|
|Non-drying clay||Claytoon , Van Aken International, CA||18150|
|Environmental chamber||I-30BLL model|