Elucidating the transcriptomic response of adult hypothalamic neuron cells to cannabidiol (CBD) exposure

Cannabidiol (CBD) actions in the brain are largely complex and not fully understood. One important brain structure that is a target for CBD, and for which health-beneficial CBD consequences can be seen is the hypothalamus. Given the fundamental role of gene expression in the hypothalamus's regulatory functions, this study undertook the analysis of changes arising in hypothalamic cells' transcriptomes following various CBD treatments. For this purpose, we used a hypothalamic cellular model, namely we employ adult-derived mHypoA-2/12 mouse cell lines. In the course of the study, the neural cells were treated with different CBD doses (ranging from 0.325 to 3 µM) and vehicle as a control for 6 and 24 hours. The experiment setup allowed us to evaluate both the dosage and time-dependent effect of CBD on hypothalamic cells, particularly on their viability, apoptosis, and transcriptome profile. Overall design: The adult mouse hypothalamic cell lines mHypoA-2/12 (Clu177) were purchased from Cedarlane (Canada) and cultured in recommended conditions. After culture stabilization, the cells were supplemented with CBD (0.325, 0.75, 1.5 and 3 µM) and cultured either for 6 hours (n = 3 for each concentration) or for 24 hours (n = 3 for each concentration). The vehicle control was 0.1% DMSO in the DMEM with GlutaMAX. When treatment time passed, cells were trypsinized, transferred to centrifuge tubes, and used for RNA isolation. Purified RNA was used for 3'mRNA-Seq to evaluate changes that arise in the transcriptome of the hypothalamic cells following different CBD treatments.