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A Behavioral Assay to Examine the Effects of Kavalactones on Caenorhabditis elegans Neuromuscular Excitability

行为分析检测卡瓦内酯对于秀丽隐杆线虫神经肌肉兴奋性的影响

BK Bwarenaba B. Kautu *
JC Jessie Chappel *
KS Kellie Steele
JP Juliana Phillips
MM M. Shawn Mengarelli

 (*contributed equally to this work) 发布: 2018年09月20日第8卷第18期 DOI: 10.21769/BioProtoc.3008 浏览次数: 5361

评审: Pengpeng LiNarayan SubramanianManish Chamoli

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Abstract

Kavalactones are a class of lactone compounds found in Kava, a traditional beverage from the South Pacific Islands that is derived from the root of Piper methysticum. When consumed, these compounds produce sedative and anxiolytic effects, suggesting their potent actions on the nervous system. Here, we provide a protocol to examine the effects of kavalactones on C. elegans neuromuscular excitability. Our methodology could provide insight into the neurophysiological actions of kavalactones.

Keywords: Kava (卡瓦) search Kavalactones (卡瓦内酯) search Lactone (内酯) search C. elegans (秀丽隐杆线虫) search Neuromuscular (神经肌肉的) search Pacific (太平洋) search Beverage (饮料) search

Background

Kava, a tranquilizing beverage from the Pacific Islands, has been consumed by Pacific Islanders for centuries (Rowe et al., 2011; Kautu et al., 2017). Kava contains a group of lipophilic compounds called kavalactones, which are believed to be responsible for the sedative, anxiolytic, and other therapeutic effects of the drink (Rowe et al., 2011; Savage et al., 2015; Kautu et al., 2017). Here, we provide a protocol to examine the effects of kavalactones on neuromuscular activity, using C. elegans as a model system (Kautu et al., 2017). In our assay, we showed that administration of aqueous kavalactone solution induced epileptic-like convulsions and paralysis in a dose-responsive manner. These manifestations are suggestive of the modulatory actions of kavalactones on the neuromuscular junction. Thus, our protocol could provide important insight into the neurophysiological actions of kavalactones.

Materials and Reagents

  1. Petri dishes, sterile (Carolina Biological Supply, catalog number: 741248 )
  2. 2 ml centrifuge tubes (Thomas Scientific, catalog number: 111572LK )
  3. P1000 pipette tips, sterile (Carolina Biological Supply, catalog number: 215060 )
  4. P200 pipette tips, sterile (Carolina Biological Supply, catalog number: 215050 )
  5. 0.5 L bottle
  6. E. coli OP50 (Carolina Biological Supply, catalog number: 155073 )
  7. C. elegans N2 (wild-type)
  8. Kava pills (Gaia Herbs, catalog number: 90A10060 )
  9. NaCl (Sigma-Aldrich, CAS: 7647-14-15)
  10. Peptone (Sigma-Aldrich, catalog number: 70176-100G )
  11. Difco Bacto agar (Carolina Biological Supply, catalog number: 156783B )
  12. CaCl2 dihydrate (Fisher Scientific, catalog number: C79 500 )
  13. MgSO4 (Sigma-Aldrich, CAS: 7487-88-9)
  14. Potassium Phosphate Monobasic (KH2PO4) (Sigma-Aldrich, CAS: 7778-77-0)
  15. Potassium Phosphate Dibasic (K2HPO4) (Sigma-Aldrich, CAS: 7758-11-4)
  16. Cholesterol (Fisher Science Education, CAS: 57-88-5)
  17. Hypochlorite solution (Sigma-Aldrich, CAS: 7681-52-9)
  18. NaOH (Sigma-Aldrich, CAS: 1310-73-2)
  19. 95% ethanol (Sigma-Aldrich, CAS: 64-47-5)
  20. 1 M CaCl2 stock solution (see Recipes)
  21. 1 M MgSO4 stock solution (see Recipes)
  22. 1 M Potassium Phosphate stock solution (pH 6) (see Recipes)
  23. 5 mg/ml Cholesterol stock (see Recipes)
  24. Nematode Growth Medium (NGM) Agar plates (see Recipes)
  25. Bleach sodium hypochlorite solution (see Recipes)
  26. Kava stock solution (5 mg/ml) (see Recipes)

Equipment

  1. P200 and P1000 micropipette
  2. Sharp Edged Scissors
  3. Autoclave
  4. Bench-top Micro Centrifuge (Oxford Lab Products, model: C12V )
  5. Meiji EMT Stereomicroscope on PBH Stand (Meiji Techno, models: EMT-1 , PBH Stan d, MA502 )
  6. Microscope Digital Camera (OMAX Microscope, catalog number: A3550UPA-R75 )
  7. Platinum wire worm pick (Genesee Scientific, catalog number: 59-AWP )
  8. Sterile incubator

Software

  1. OMAX ToupView 3.7
  2. Microsoft Office 2010 Excel (Microsoft Corporation, Redmond, USA)

Procedure

English
中文翻译

文章信息

版权信息

© 2018 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Kautu, B. B., Chappel, J., Steele, K., Phillips, J. and Mengarelli, M. S. (2018). A Behavioral Assay to Examine the Effects of Kavalactones on Caenorhabditis elegans Neuromuscular Excitability. Bio-protocol 8(18): e3008. DOI: 10.21769/BioProtoc.3008.

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分类

神经科学 > 行为神经科学 > 实验动物模型

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