Abstract
BackgroundWhole-plant cannabis extracts are consumed by the public for medical and non-medical (“recreational”) purposes but are poorly researched compared to pure cannabinoids. There is emerging evidence that cannabis extracts comprising complex mixtures of cannabinoids may have different biological effects from that of pure cannabinoids. In the current study, we sought to assess the effect of whole-plant cannabis extracts produced from different chemotypes of cannabis on the normal behavior of zebrafish larvae.MethodsThree cannabis plant chemotypes were used in this study that contained either high amounts of THC, high amounts of CBD, high but equal amounts of THC and CBD, or low but equal amounts of THC and CBD. Following solvent extraction, liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) was performed for the detection and quantitation of target cannabinoids. Larval zebrafish behavioral models were subsequently used to assess the effect of the four different whole-plant cannabis extracts on the normal larval behavior using the DanioVision behavioral tracking systems and software. To compare, changes in the behavior activity levels for 30 min periods were compared to controls using 2-way ANOVA with multiple comparisons followed by a Bonferroni post hoc test.ResultsIt was found that the whole-plant extracts that contained high levels of THC had similar effects on larval behavior, while the high CBD and low THC:CBD extracts produced distinct effects on normal larval behavior. Exposure of larvae to concentration-matched levels of THC and CBD found in the extracts revealed that a subset of the cannabis extracts tested had similar behavioral profiles to the pure cannabinoids while others did not.ConclusionsTo our knowledge, this is the first study to test and compare the bioactivity of different whole-plant cannabis extracts in larval zebrafish. This work will provide a framework for future studies of distinct cannabis extracts and will be useful for comparing the bioactivity of extracts from different cannabis chemotypes as well as extracts made through various heating processes. It will also act as the first stage of assessment before testing the extracts against zebrafish models of toxicity and disease.
Highlights
Whole-plant cannabis extracts are consumed by the public for medical and non-medical (“recrea‐ tional”) purposes but are poorly researched compared to pure cannabinoids
An entourage effect has been described between ∆-9 tetrahydrocannabinol (THC) and cannabidiol (CBD), the main cannabinoids found in cannabis (Russo & Guy, 2006; Russo, 2011)
We sought to assess the effect of whole-plant cannabis extracts produced from different chemotypes of cannabis on the normal behavior of zebrafish larvae
Summary
Whole-plant cannabis extracts are consumed by the public for medical and non-medical (“recrea‐ tional”) purposes but are poorly researched compared to pure cannabinoids. An entourage effect has been described between ∆-9 tetrahydrocannabinol (THC) and cannabidiol (CBD), the main cannabinoids found in cannabis (Russo & Guy, 2006; Russo, 2011) These interactions are complex and may result in the inhibition of their individual effects depending on a number of factors, such as the ratios or levels of each compound and whether the compounds are administered sequentially or simultaneously (Russo, 2011; Canada Go, 2018; Zuardi et al, 2012; Davis & Hatoum, 1983; Freeman et al, 2019a). Several of the models developed use larval behavior as a platform to assess the activity of cannabinoids (Akhtar et al, 2013; BoaAmponsem et al, 2019; Ellis et al, 2018; Hasumi et al, 2020; Samarut et al, 2019; Chousidis et al, 2020; Carty et al, 2019). The light/ dark preference test is based on larvae exhibiting an innate avoidance response to dark environments and is considered an anxiety-like behavior that can be used to assess the efficacy of anxiolytics (Maximino et al, 2010; Steenbergen et al, 2011)
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