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CB1 BUT NOT CB2 CANNABINOID RECEPTOR INCREASES NEURITE EXTENSION IN HUMAN NEUROBLASTOMA
Author(s) -
Lyons Erica Lynn,
Kabler Sandra,
Howlett Allyn,
Kovach Alexander L.,
Thomas Brian
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.540.2
Subject(s) - cannabinoid receptor , neurite , cannabinoid , receptor , cannabinoid receptor type 2 , endocannabinoid system , gpr18 , transfection , 2 arachidonoylglycerol , stimulation , chemistry , microbiology and biotechnology , neuroblastoma , biology , medicine , endocrinology , cell culture , biochemistry , gene , genetics , in vitro , agonist
Despite neurodegenerative disease affecting more than five million Americans, there is not yet sufficient understanding of the underlying mechanism of disease to cure or even halt progression. Transplanted neurons may represent a potential treatment if they could successfully integrate with host brain tissue. Both CB1 and CB2 receptors are located in neuronal growth cone membranes, implying that cannabinoid signaling may play a role in neurite extension or guidance (Harkany et al. 2007). CB 1 cannabinoid signaling has been shown to be active in neuronal extension, yet the differences between CB 1 R and CB 2 R signaling remain unclear (Berghuis et al. 2007, Duff et al. 2013). We have created a novel stable overexpression model to investigate the role of cannabinoid signaling in human neuronal extension. We quantified the endogenous production of cannabinoid ligand 2‐Arachidonoyl Glycerol (2‐AG) and its isomer 1‐AG in SHSY5Y human neuroblastoma at an abundance of 4,000 nanograms Total AG per million cells. This AG was likely 2‐AG produced by DAGL enzyme activity because calcium stimulation by Oxotremorine M increased Total AG abundance to 6,000 nanograms per million cells while inhibition of DAGL by THL decreased Total AG abundance to less than 100 nanograms per million cells. We transfected SHSY5Y cells with either CB1 or CB2 receptors, purified and maintained the transfection using geneticin selection, and isolated clonal cell lines that stably overexpress either CB 1 R (CB1XS) or CB 2 R (CB2XS). Stable overexpression of cannabinoid receptors did not alter mRNA abundance of the enzymes that synthesize and degrade the endogenous cannabinoids 2‐AG and AEA. Stable overexpression of CB 1 R but not CB 2 R increased the extension length from 30 to 200 micrometers per neurite. Our research suggests a role for CB 1 R but not CB 2 R mediated cannabinoid signaling in neurite development that may provide insights in development of therapies that support functional integration of transplanted neurons into host tissue. Support or Funding Information Support from NIDA grant DA03690 and T32 grant HL091797 is appreciated. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .