Giorgi, Victoria et al. (2021) International Worm Meeting "C. elegans as a model for studying Cannabis sativa extracts"

Garcia, Carlos, Carrera, Ines, Fernandez, Santiago, Castro, Rosina, Giorgi, Victoria, Vila, Agustina, Pazos, Mariana

[International Worm Meeting, 2021]

The Cannabis genus includes a variety of plants that are a valuable source of secondary bioactive metabolites, including flavonoids, terpenes, and cannabinoids. These metabolites are responsible for a wide variety of biological activities such as anti-inflammatory, anti-oxidant, psychotropic and others. The long term objective of this project is to develop C. elegans as a model for studying the bioactive potential of different extract types from cannabis plants. Previous studies showed that aqueous extracts from Cannabis sativa leaves exhibit nematicidal activity on plant parasitic nematodes, and suggested that phytocannabinoids present in the extract are responsible for this effect. However, little is known about the nematicidal activity from extracts of other parts of cannabis plants, including female flowers where phytocannabinoids accumulate in highest amounts. C.elegans has proven to be an excellent model for the discovery of new anthelmintic drugs and its mechanism of action. In this sense extracts from flowers of two Cannabis plant varieties which differ in its ratio of tetrahydrocannabinolic acid (THCA)/ cannabidiolic acid (CBDA) content were prepared and its activity on C. elegans motility assessed. The phytocannabinoids profile of the extracts was studied and the main phytocannabinoids were identified and quantified by HPLC-DAD. Automated motility assays were conducted with N2 worms at 10 microM and 100 microM in the main cannabinoid (CBDA or THCA). Failure to see a motility effect in this background prompted us to use mutants with a compromised cuticle. C.elegans has a robust cuticle that functions as a physical barrier against chemicals which may represents a problem for compounds testing, such as acidic phytocannabinoids. In this sense, motility assays were performed using mutants in the bus-8 gene, which encodes a predicted glycosyltransferase required for cuticle integrity. Cannabis flower extracts caused severe reduction in motility in bus-8 mutants at 50 microM and 100 microM in the main cannabinoid. We are currently analyzing purified compounds to dissect the complex effect of these extracts and using mutant analysis to understand its mechanism of action.

Mariana Simaes et al. (2006) European Worm Meeting "Phenotypic Analysis and Mapping of a New C. elegans Mab Mutant, mab-29"

Mariana Simaes, Alison Woollard

[European Worm Meeting, 2006]

Mariana Simes and Alison Woollard. Mab (Male ABnormal) mutants have defects in male tail copulatory structures and therefore have a reduced ability to mate. Nonetheless, these mutations can be isolated without difficulties accompanying lethality since the normal population consists of hermaphrodites. In a recent F1 screen for new Mab mutants, one such mutant was isolated, mab-29. Male tails have swollen bursa/ rays and reduced fan, suggesting a defect in morphogenesis. Extensive phenotypic analysis of the male tail using conventional microscopy and specific tissue markers shows abnormal attachment of the rays and phasmid to the hypodermal sheath, resulting in the striking appearance of the lumpy and amorphous rays and reduced fan.. By using recombination mapping, mab-29 has been mapped to an interval on Linkage Group X. We are using a combination of three-factor crosses and SNP snip-mapping approaches to narrow the genomic region in which mab-29 is found. Snip mapping is being carried out using snips in the interval of about three map units, between lin-2 and unc-9.. Our mapping strategy and progress towards defining the mab-29 map position will be presented. Further analysis depends on the nature of this gene.