Transcriptomic and Proteomic Analyses of Mouse Cerebellum Reveals Alterations in RasGRF1 Expression Following In Vivo Chronic Treatment with Delta 9-Tetrahydrocannabinol

Abstract

We have applied transcriptomic and proteomic techniques to identify changes in the RNA and the protein levels in the mouse cerebellum after chronic treatment with Delta(9)-tetrahydrocannabinol (THC). Among approximately 14,000 transcripts in a mouse cDNA microarray library, we found 11 genes with altered expression. RasGRF1, a neuron-specific Ras guanine nucleotide exchange factor, showed a reduction both at the RNA and protein levels with a specific decrease of the protein pool associated to cell membranes. In addition, proteomic analysis on cerebellum obtained from chronically THC-treated mice detected quantitative changes of additional 27 spots, mostly in the membranous fraction. We found enrichment of alpha (Galphao, Galphaq) and beta subunits (beta4/beta2 and beta5) of guanine nucleotide-binding proteins and of two calcium-binding proteins, calretinin and hippocalcin-like protein-1. In addition, we also detected a significant increase in the membrane fraction of proteins involved in exo-endocytosis such as septins, dynamin-1, and vesicle protein sorting 29. By western blotting, we confirmed increased membrane localization of calretinin and of dynamin-1 isoforms with higher isoelectric point, indicative for an underphosphorylated state of the molecule. In conclusion, our results indicate that chronic THC modulates the expression and subcellular localization of proteins implicated in Ras signaling, calcium-buffering potential, and trafficking.

Characterization of the Endocannabinoid System in Human Neuronal Cells and Proteomic Analysis of Anandamide-induced Apoptosis

“…This is the first proteomic approach to the understanding of AEA pro-apoptotic activity. By the way, the first ever proteomic analysis on the effects of the cannabis-derived ⌬ 9 -tetrahydrocannabinol in chronic users (45), and in mouse cerebellum (46), appeared during the preparation of this manuscript.…”

Genes and Pathways Co-associated with the Exposure to Multiple Drugs of Abuse, Including Alcohol, Amphetamine/Methamphetamine, Cocaine, Marijuana, Morphine, and/or Nicotine: a Review of Proteomics Analyses

“…Of these proteins, ten were suppressed by THC and only one was induced, which might indicate a reduced synaptic efficiency in the PFC caused by THC treatment. Via proteomics analysis, Colombo et al [49] identified 24 unique differentially expressed proteins in the cerebellum of chronically THC-treated mice. Among these proteins, several alpha and beta subunits of guanine nucleotidebinding proteins, calcium-binding proteins (calretinin and hippocalcin-like protein 1), and dynamin-1 had induced expression.…”

Endocannabinoids Stimulate Human Melanogenesis via Type-1 Cannabinoid Receptor

“…These data indicate that both CB subtypes were functional to a similar extent in human NHEM cells (Table 1). However, some discrepancies were observed between the activity and mRNA expression of AEA-and 2-AG-metabolizing enzymes, a finding that is not unprecedented for ECS elements (35,44). Overall, the present data demonstrate that primary human melanocytes have a complete and fully functional ECS.…”

“…Overall, the present data demonstrate that primary human melanocytes have a complete and fully functional ECS. Induction of Apoptosis-AEA at micromolar concentrations has been shown to trigger apoptosis in different cell types (34,44,45). Here we treated NHEM cells for 24 h with different amounts of mAEA, a non-hydrolyzable analog that binds the same targets and with the same affinity as AEA (3) and shows the same functional effects as the natural compound (3,4).…”