<meta charset="utf8"></head><body>秀丽隐杆线虫中挥发性信息素反应的提取和定量

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>10 cm Petri dishes</td>
			<td>Falcon</td>
			<td>25373-100</td>
			<td>Falcon bacteriological Petri dish 100 x 15 mm</td>
		</tr>
		<tr>
			<td>6 cm Petri dishes</td>
			<td>Falcon</td>
			<td>25373-085</td>
			<td>Falcon bacteriological Petri dish 60 x 15 mm</td>
		</tr>
		<tr>
			<td>C. remanei (EM464)</td>
			<td>CGC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Eppendorf</td>
			<td>centrifuge 5418</td>
			<td>&#160;Any brand should work.</td>
		</tr>
		<tr>
			<td>Chemoattraction assay plates</td>
			<td>Homemade solution</td>
			<td>N/A</td>
			<td>1.5% agar, 25 mM NaCl, 1.5 mM Tris-base, and 3.5 mM Tris-Cl</td>
		</tr>
		<tr>
			<td>Cholesterol</td>
			<td>Alfa Aesar</td>
			<td>57-88-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting Microscope</td>
			<td>Leica</td>
			<td>LeicaMZ75</td>
			<td>&#160;Any brand should work.</td>
		</tr>
		<tr>
			<td>E. Coli OP50</td>
			<td>CGC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Koptec</td>
			<td>64-17-5</td>
			<td></td>
		</tr>
		<tr>
			<td>fog-2(q71) (JK574)</td>
			<td>CGC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>him-5(e1490)(CB4088)</td>
			<td>CGC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Household bleach</td>
			<td>Clorox Germicidal bleach concentrated</td>
			<td></td>
			<td>&#160;Bleach</td>
		</tr>
		<tr>
			<td>M9 buffer</td>
			<td>Homemade solution</td>
			<td>N/A</td>
			<td>3 g KH2PO4, 11.3 g Na2HPO4.7H2O, 5 g NaCl, H2O to 1 L. Sterilize by autoclaving. Add 1 mL 1 M MgSO4 after cool down to room temperature.</td>
		</tr>
		<tr>
			<td>Magnesium Sulfate, Anhydrous, Powder</td>
			<td>Macron</td>
			<td>M1063-500GM-EA</td>
			<td></td>
		</tr>
		<tr>
			<td>Microwave</td>
			<td>TOSHIBA</td>
			<td>N/A</td>
			<td>&#160;Any brand should work.</td>
		</tr>
		<tr>
			<td>N2</td>
			<td>CGC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Sigma-aldrich</td>
			<td>S318-3</td>
			<td>1 M</td>
		</tr>
		<tr>
			<td>NGM plates solution</td>
			<td>Homemade solution</td>
			<td>N/A</td>
			<td>2.5 g Peptone, 18 g agar, 3 g NaCl, H2O to 1 L.Sterilize by autoclaving. Once the autoclave is done (2 h), wait until the temperature of the medium drops to 65 &#176;C. Put on a hotplate at 65 &#176;C and stir. Then add the following, waiting 5 min between each to avoid crystallization: 1 mL CaCl2 (1 M), 1 mL MgSO4 (1 M), 25 mL K3PO4 (1 M, pH=6), 1 mL Cholesterol ( 5 mg/mL in ethanol).</td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>Bemis</td>
			<td>13-374-10</td>
			<td>Bemis Parafilm M Laboratory Wrapping Film</td>
		</tr>
		<tr>
			<td>Peptone</td>
			<td>VWR</td>
			<td>97063-324</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipet- aid</td>
			<td>Drummond Scientific&#160;</td>
			<td>4-000-100</td>
			<td>&#160;Any brand should work.</td>
		</tr>
		<tr>
			<td>Plastic paper&#160;</td>
			<td>Octago</td>
			<td>Waterproof Screen Printing Inkjet Transparency Film</td>
			<td>https://www.amazon.com/Octago-Waterproof-Transparency-Printing-Printers/dp/B08HJQWFGD</td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Sigma-aldrich</td>
			<td>SLBP2366V</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium phosphate</td>
			<td>Spectrum</td>
			<td>7778-77-0</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette</td>
			<td>Eppendorf</td>
			<td>SKU: EPPR4331; MFG#: 2231300006</td>
			<td>20 - 200 &#181;L, 100 - 1000 &#181;L, any brand should work.</td>
		</tr>
		<tr>
			<td>Rotator</td>
			<td>Labnet</td>
			<td>SKU: LI-H5500</td>
			<td>&#160;Labnet H5500 Mini LabRoller with Dual Direction Rotator. Any brand should work.</td>
		</tr>
		<tr>
			<td>Sodium chloride</td>
			<td>VWR</td>
			<td>7647-14-5</td>
			<td></td>
		</tr>
		<tr>
			<td>sodium phosphate dibasic</td>
			<td>Sigma-aldrich</td>
			<td>SLCG3888</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-base</td>
			<td>Sigma-aldrich</td>
			<td>77-86-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Tris-Cl&#160;</td>
			<td>Roche</td>
			<td>1185-53-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptone</td>
			<td>VWR</td>
			<td>97063-390</td>
			<td></td>
		</tr>
		<tr>
			<td>Vortex</td>
			<td>Scientific industries</td>
			<td>Vortex-Genie 2</td>
			<td>&#160;Any brand should work.</td>
		</tr>
		<tr>
			<td>WormLab system&#160;</td>
			<td>MBF Bioscience</td>
			<td>N/A</td>
			<td>https://www.mbfbioscience.com/help/WormLab/Content/home.htm; https://www.mbfbioscience.com/products/wormlab/</td>
		</tr>
		<tr>
			<td>Wormpicker</td>
			<td>Homemade&#160;</td>
			<td>N/A</td>
			<td>made with platinum and glass pipet tips</td>
		</tr>
	</tbody>
</table>

volatile sex pheromone extraction and chemoattraction assay in caenorhabditis elegans

The interplay between chemical communication and reproductive success is a fundamental principle across the animal kingdom1,2,3,4,5,6,7,8,9,10. Sex pheromones trigger a wide array of sexually dimorphic behaviors essential for locating mates, coordinating the steps involved in finding and attracting a partner, and ultimately promoting the propagation of a species11,12,13,14,15,16,17. Significant progress has been made in understanding pheromone signaling, but the molecular mechanisms, neural circuits, and computational processes governing these interactions often remain incompletely defined18,19,20,21,22,23,24,25,26.
The nematode Caenorhabditis elegans provides a powerful model for dissecting these questions. Notably, C. elegans exhibits an unusual reproductive strategy-hermaphrodites can self-fertilize but also outcross with males27,28,29,30,31,32,33. This flexibility requires a robust communication system to signal reproductive status. C. elegans is known for its well-characterized water-soluble pheromones, the ascarosides, which play varied roles in development, behavior, and social interactions. Recent discoveries have unveiled a distinct class of volatile sex pheromones employed by nematodes. These pheromones are specifically produced by sexually mature C. elegans and C. remanei virgin females and sperm-depleted hermaphrodites, serving as an attractant for adult males29,34,35. This attractant exhibits remarkable sexual dimorphism in its production and perception. The female somatic gonad governs the synthesis of this volatile sex pheromone, and production dynamically reflects reproductive status, ceasing upon mating and resuming several hours later29,34.
Understanding nematode sex pheromone communication provides insights into the evolution of chemical communication systems, the interplay between reproductive state and behavior, and the mechanisms underlying sexually dimorphic neural processing24,26,36,37,38,39. Studies implicate the amphid neuron AWA in males as critical for pheromone detection, with the G-protein-coupled receptor SRD-1 playing a key role in pheromone detection in males24. C. elegans is well-suited for studying animal chemical communication, especially sex pheromone signaling, due to its reliance on the olfactory system for mate-searching. While much is known about ascaroside signaling, the volatile sex pheromone system offers unique opportunities for comparison25,26,36,40,41,42,43,44,45,46,47,48,49,50, 
51,52,53,54,55,56,57. Moreover, C. elegans is a powerful genetic model organism due to its fully sequenced genome, clearly defined cellular lineage, and well-characterized olfactory mutants.
However, the complete neural circuitry involved in processing this pheromone, the computations that translate its perception into targeted mate-searching behaviors, and its biosynthesis regulation remain to be fully elucidated. Further investigations into these processes are crucial for understanding the diverse mechanisms governing animal communication and reproductive behaviors. The identification of key genes involved in pheromone synthesis, secretion, and perception promises to unveil novel molecular players in animal communication.&#160;The assays&#160;described here provide a basis to address these&#160;questions.

<meta charset="utf8"></head><body>作者聚焦：检查挥发性信息素对雄性 秀丽隐杆线 虫行为的影响

秀丽隐杆线虫中的挥发性信息素提取和趋化试验

Chemical communication is vital in organismal health, reproduction, and overall well-being. Understanding the molecular pathways, neural processes, and ...

volatile-sex-pheromone-extraction-chemoattraction-assay

Research

JoVE Journal

Behavior

Volatile Sex Pheromone Extraction and Chemoattraction Assay in Caenorhabditis elegans

1.2K 次观看.  California Institute of Technology. 我们的研究使用趋化性测定法检查了挥发性信息素如何影响雄性秀丽隐杆线虫的行为。它旨在确定信息素产生的分子机制，其神经元机制是感知。响应信息素的个体蠕虫间变异性，再加上环境因素（如温度、湿度和光线）的影响，可能会在数据中引入大量噪声。此外，蠕虫行为的复杂性使数据解释变得复杂。应对这些挑战需要仔细的实验设计。...

视频: 作者聚焦：检查挥发性信息素对雄性 秀丽隐杆线 虫行为的影响

Chemical communication is vital in organismal health, reproduction, and overall well-being. Understanding the molecular pathways, neural processes, and computations governing these signals remains an active area of research. The nematode Caenorhabditis elegans provides a powerful model for studying these processes as it produces a volatile sex pheromone. This pheromone is synthesized by virgin females or sperm-depleted hermaphrodites and serves as an attractant for males.
This protocol describes a detailed method for isolating the volatile sex pheromone from several C. elegans strains (WT strain N2, daf-22, and fog-2) and C. remanei. We also provide a protocol for quantifying the male chemotaxis response to the volatile sex pheromone. Our analysis utilizes measurements such as chemotaxis index (C.I.), arrival time (A.T.), and a trajectory plot to compare male responses under various conditions accurately. This method allows for robust comparisons between males of different genetic backgrounds or developmental stages. Furthermore, the experimental setup outlined here is adaptable to investigating other chemoattraction chemicals.

This protocol establishes methods for extracting and quantifying responses to the volatile sex pheromone in C. elegans, providing tools to study chemical communication and navigation trajectory.

作者聚焦：检查挥发性信息素对雄性 秀丽隐杆线 虫行为的影响

作者聚焦：检查挥发性信息素对雄性秀丽隐杆线虫行为的影响