Delta-9 tetrahydrocannabinol (THC) effects on the cortisol stress response in bovine granulosa cells

Dufour, J.; Sabry, Reem; Khokhar, Jibran Y.; Favetta, Laura A.

doi:10.1016/j.tiv.2022.105549

Cited by 1 publication

(4 citation statements)

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“…Together, DNMTs and TETs/TDG establish, maintain and erase CpG methyl marks, contributing to overall gene expression [ 90 ]. The detected increase in DNMT1 mRNA and protein expression may be indicative of increased cell replication, but previous research in our lab found no significant changes in cell counts following THC exposure [ 83 ]. Cells might be increasing DNMT1 transcripts to compensate for reduced 5-mC marks following THC exposure.…”

Section: Discussionmentioning

confidence: 76%

“…These values were additionally reported by Whan et al [ 82 ] as being representative of blood plasma concentrations found in therapeutic and recreational cannabis users. THC concentrations chosen for this study are also reflective of previously published research from our laboratory investigating THC on sperm parameters [ 62 ], GC viability [ 83 ], and oocyte and embryo quality [ 54 ].…”

Section: Discussionmentioning

confidence: 99%

“…Previously in our laboratory, GCs exposed to therapeutic and recreational doses of THC did not display compromised cell viability or apoptosis [ 83 ]. Misner et al [ 54 ] performed a transcriptome analysis which revealed that THC was dysregulating transcript expression in COCs.…”

Section: Discussionmentioning

confidence: 99%

“…In this investigation, GCs are used as they are an excellent indicator of oocyte health and crucial to oocyte development by providing a suitable microenvironment during oogenesis [ 74 – 81 ]. THC concentrations were chosen based on THC plasma concentrations detected in recreational users and after therapeutic use [ 82 ] and are in line with previous research in our laboratory that assessed THC’s effects in gametes and blastocysts [ 54 ], and GCs [ 83 ]. In addition, THC was detected in follicular fluid of ART patients at a concentration of 0.03243 μM, which is the range of the therapeutic dose (0.032 μM, [THC]) utilized in our study [ 57 ].…”

Section: Introductionmentioning

confidence: 99%

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DNA methylation, but not microRNA expression, is affected by in vitro THC exposure in bovine granulosa cells

Floccari,

Sabry,

Choux

et al. 2024

BMC Pharmacol Toxicol

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Background A global increase in cannabis use has led to questions about its effects on fertility. The rise in consumption amongst women of reproductive age is a growing concern, as this group is vulnerable in terms of reproductive health. Ample evidence suggests that the psychoactive component of cannabis, Δ9-Tetrahydrocannabinol (THC), interacts with the endocannabinoid system (ECS), that helps regulate mammalian reproduction. This study aimed to research the epigenetic effects of THC in bovine granulosa cells (GCs) by (1) investigating global DNA methylation via measuring 5-mC and 5-hmC levels; (2) measuring key methylation regulators, including the methylating enzymes DNMT1, DNMT3a, DNMT3b and the demethylases TDG and TET1/2/3; and (3) assessing fertility-associated miRNAs key in developmental competency, including miR-21, -155, -33b, -324 and -346. Methods Bovine GCs were used as a translational model for reproductive toxicity in humans. To determine THC effects, GCs were isolated from Cumulus-Oocyte-Complexes (COCs) from bovine ovaries, cultured in vitro for 7 days, or until confluent, and cryopreserved at passage 1 (P1). For experimentation, cells were thawed, cultured until passage 2 (P2), serum restricted for 24-h and treated for 24-h in one of five groups: control, vehicle (1:1:18 ethanol: tween: saline) and three clinically relevant THC doses (0.032, 0.32 and 3.2 μM). Global methylation was assessed by measuring 5-mC and 5-hmC levels with flow cytometry. To assess mRNA and protein expression of methylation regulators and miRNA profiles, qPCR and Western Blotting were utilized. Shapiro-Wilk test was used to determine normality within datasets. One-way ANOVA was applied to determine statistical significance using GraphPad Prism 6.0.0. Results Results indicate a significant decrease (p = 0.0435) in 5-mC levels following low THC exposure, while no changes were observed in 5-hmC levels. A significant increase in DNMT1 following high THC exposure at the RNA level (p < 0.05) and a significant increase following low THC exposure at the protein level (p = 0.0048) were also observed. No significant differences were observed in DNMT3a/3b, TDG, TET1/2/3 mRNAs or in any of the miRNAs analyzed. Conclusions This research suggests that THC mainly affects DNA methylation, but not miRNA profiles, ultimately altering gene expression and likely impairing oocyte competence, maturation, and fertilization potential.

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Section: Discussionmentioning

confidence: 76%