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

Bwarenaba B. Kautu; Jessie Chappel; Kellie Steele; Juliana Phillips; M. Shawn Mengarelli

doi:10.21769/BioProtoc.3008

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Peer-reviewed

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 ^* email

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: 4748

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)

Kavalactones (卡瓦内酯)

Lactone (内酯)

C. elegans (秀丽隐杆线虫)

Neuromuscular (神经肌肉的)

Pacific (太平洋)

Beverage (饮料)

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

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

Equipment

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

Software

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

Procedure

English

中文翻译

文章信息

版权信息

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.