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Role of cell wall proteins in Saccharomyces cerevisiae’s response to commercial formulations of glyphosate and contribution of Ty elements to adaptation mechanisms

ABSTRACT: The use of glyphosate-based herbicides is widespread, despite its extensive use its effects are yet to be deciphered completely. The additives in commercial formulations of glyphosate though labelled inert when used individually, has adverse effects when used in combination with other additives and the active ingredient. An RNA seq study was performed to thoroughly investigate the changes that occur in a sensitive and a resistant strain at the transcriptome level on exposure to a commercial formulation of glyphosate. Changes in gene expression of genes contributing to numerous pathways involved in crucial cellular functions such as DNA replication, MAPK signaling, meiosis, cell wall synthesis etc. were found. Common fragile sites were found to play a role in adaptation mechanisms used by cells to attain resistance on long term exposure to a CFG. The cell wall structure acts as a protective barrier in alleviating the stress caused by exposure to Cr41, the thicker the cell wall the more resistant the cell is against CFG. Hence, a detailed study of the changes occurring at the genome and transcriptome level Is essential to better understand the possible effects of CFG on the cell as a whole.

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE135473 | GEO | 2020/04/29

REPOSITORIES: GEO

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Project description:Glyphosate is one of the most widely used herbicides globally. It acts by inhibiting an enzyme in an aromatic amino synthesis pathway specific to plants and microbes, leading to view that glyphosate poses no risk to other organisms. However, there is growing concern that glyphosate is associated with detrimental health effects in humans, and an ever-increasing body of evidence suggests that glyphosate affects other animals including pollinating insects such as bees. Although pesticides have long been considered a contributing factor in the decline of wild bee populations most research on bees has focussed on demonstrating and understanding the effects (particularly sublethal ones) of insecticides. To assess whether glyphosate poses a potential risk to bees we characterised the changes in survival, behaviour, digestive tract proteome and microbiome in the bumblebee Bombus terrestris after chronic exposure to field relevant doses of glyphosate alone and as part of the commercially available product RoundUp Optima+®. Regardless of source, changes in response to herbicide exposure in important cellular and physiological processes in the digestive tract of B. terrestris were observed, with the abundances of proteins associated with oxidative stress regulation, metabolism, cellular adhesion, the extracellular matrix, and various signalling pathways being altered. Interestingly, endocytosis, oxidative phosphorylation, the TCA cycle, and carbohydrate, lipid, and amino acid metabolism were differentially altered depending on whether the exposure source was glyphosate AI or RoundUp Optima+®. In addition, RoundUp Optima+®, but not the active ingredient glyphosate, impacted fungal diversity in the digestive tract microbiota. Our research provides new insights into the potential mode of action and consequences of glyphosate exposure at the molecular and cellular levels in bumblebees and highlights issues with current regulatory measures involving commercial formulations of pesticides where the impact of the co-formulants on non-target organisms are generally overlooked.

Project description:Background: Herbicides are environmental contaminants that have gained much attention due to the potential hazards they pose to human health. Glyphosate, the active ingredient in many commercial herbicides, is the most heavily applied herbicide worldwide. The recent rise in glyphosate application to corn and soy crops correlates positively with increased death rates due to Alzheimer's disease and other neurodegenerative disorders. Glyphosate has been shown to cross the blood-brain barrier in in vitro models, but has yet to be verified in vivo. Additionally, reports have shown that glyphosate exposure increases pro-inflammatory cytokines in blood plasma, particularly TNFα. Methods: Here, we examined whether glyphosate infiltrates the brain and elevates TNFα levels in 4-month-old C57BL/6J mice. Mice received either 125, 250, or 500 mg/kg/day of glyphosate, or a vehicle via oral gavage for 14 days. Urine, plasma, and brain samples were collected on the final day of dosing for analysis via UPLC-MS and ELISAs. Primary cortical neurons were derived from amyloidogenic APP/PS1 pups to evaluate in vitro changes in Aβ40-42 burden and cytotoxicity. RNA sequencing was performed on C57BL/6J brain samples to determine changes in the transcriptome. Results: Our analysis revealed that glyphosate infiltrated the brain in a dose-dependent manner and upregulated TNFα in both plasma and brain tissue post-exposure. Notably, glyphosate measures correlated positively with TNFα levels. Glyphosate exposure in APP/PS1 primary cortical neurons increases levels of soluble Aβ40-42 and cytotoxicity. RNAseq revealed over 200 differentially expressed genes in a dose-dependent manner and cell-type-specific deconvolution analysis showed enrichment of key biological processes in oligodendrocytes including myelination, axon ensheathment, glial cell development, and oligodendrocyte development. Conclusions: Collectively, these results show for the first time that glyphosate infiltrates the brain, elevates both the expression of TNFα and soluble Aβ, and disrupts the transcriptome in a dose-dependent manner, suggesting that exposure to this herbicide may have detrimental outcomes regarding the health of the general population.

Project description:Glyphosate is the herbicide most used worldwide, with no historic comparison. It is used for genetically modified crops, and particularly in Florida, it is used as a sugar cane ripener. It is also formulated to treat aquatic weeds in waterbodies as Rodeo®. Because of its extended use, it can run off or be sprayed directly into waterbodies and aquatic wildlife can be chronically exposed. Exposure in animal models has been associated with kidney and liver damage and it has been suggested as an endocrine disruptor. We exposed adult male largemouth bass for 21 days to two doses of glyphosate and Rodeo (chemically equivalent concentration of glyphosate) including 0.5 mg/L and 10 mg/L and to clean water as a control (n=4/tank in quadruplicate). Concentrations during the experiment were corroborated with LC-MS/MS. Total RNA was isolated from the trunk kidney and head kidney. RNA-seq was performed for the high doses compared to controls (n=4/treatment). Transcripts were analyzed with fish and mammalian pathway analyses. Transcripts mapped to Zebrafish metabolic pathways using Paintomics showed the most significantly enriched pathway was steroid hormone biosynthesis in the trunk kidney. Steroid hormones were measured in plasma by LC-MS/MS. Total androgens were significantly reduced at 0.5 mg/L of glyphosate and Rodeo compared to control. While 11-ketotestosterone and estrone concentration was significantly reduced in all doses. Our results show a possible down regulation of the conversion of testosterone to 11-ketotestosterone by glyphosate and Rodeo at concentrations that are allowed in U.S drinking water. Using the mammalian pathway analysis algorithm, cellular processes associated with T-cell activation/development and intracellular pH were significantly enriched in the trunk kidney after glyphosate and Rodeo exposure. Glyphosate and Rodeo produce endocrine disruption corroborated at the hormone and gene expression level. Rodeo and glyphosate share gene expression pathways, however, Rodeo had more pronounced effects in largemouth bass.

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Role of cell wall proteins in Saccharomyces cerevisiae’s response to commercial formulations of glyphosate and contribution of Ty elements to adaptation mechanisms

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