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Neuronal Nitric Oxide Synthase Critically Regulates the Endocannabinoid Pathway in the Murine Cerebellum During Development

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Abstract

The cerebellum is a major site of endocannabinoid (eCB) production and signaling. The predominant eCB within the cerebellum, 2-arachidonoylglycerol (2-AG), is produced by a metabotropic glutamate receptor type 1 (mGluR1)-initiated signaling cascade within Purkinje neurons (PNs). 2-AG retrogradely stimulates cannabinoid 1 receptors (CB1Rs) located on presynaptic terminals. The activated CB1R decreases neurotransmitter release and leads to the production of nitric oxide (NO), a gaseous molecule. Recently, our group discovered that during development in mice lacking neuronal nitric oxide synthase (nNOS−/−), PNs display an excitotoxic phenotype associated with overactivated mGluR1. Considering the importance of mGluR1 in 2-AG synthesis, the present study explored the role of nNOS-derived NO in regulating the eCB pathway within the cerebella of wildtype (WT) and nNOS−/− mice at postnatal day 7 (PD7), 2 weeks (2 W), and 7 weeks (7 W). Our analysis showed that diacylglycerol lipase α, the enzyme that catalyzes 2-AG production, was elevated at early postnatal ages, and followed by elevated levels of 2-AG in nNOS−/− cerebella compared to WT. CB1R expression in nNOS−/− cerebella was upregulated at PD7 but decreased at 2 W and 7 W when compared to age-matched WT mice cerebella. Importantly, treating organotypic nNOS−/− cerebellar slice cultures with an NO-donor–attenuated CB1R levels after 7 days in vitro. In addition, expression of the eCB hydrolases fatty acid amide hydrolase and monoacylglycerol lipase were significantly downregulated in nNOS−/− cerebella compared to WT cerebella at 7 W. Together, these results reveal a novel role for nNOS/NO in regulating eCB signaling in the cerebellum.

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Funding

This study was supported by a CIHR grant (MOP-133504) to W-Y L. VT and MJEM have been awarded Ontario Graduate Scholarships.

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Authors and Affiliations

  1. Graduate Program of Neuroscience, The University of Western Ontario, London, N6A 5B7, Canada

    Vasiliki Tellios, Matthew J. E. Maksoud & Wei-Yang Lu

  2. Robarts Research Institute, London, N6A 5B7, Canada

    Vasiliki Tellios, Matthew J. E. Maksoud, Ravneet Nagra & Wei-Yang Lu

  3. Department of Physiology and Pharmacology, The University of Western Ontario, London, N6A 5B7, Canada

    Gurneet Jassal & Wei-Yang Lu

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Tellios, V., Maksoud, M.J.E., Nagra, R. et al. Neuronal Nitric Oxide Synthase Critically Regulates the Endocannabinoid Pathway in the Murine Cerebellum During Development. Cerebellum 22, 1200–1215 (2023). https://doi.org/10.1007/s12311-022-01493-2

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