Effect of N-arachidonylethanolamine on axon regeneration of the dorsal root ganglion

doi:10.12307/2022.405

Abstract

Abstract: BACKGROUND: Nerve axon regeneration is mainly affected by its own regenerative ability and inhibitory external environment. N-arachidonylethanolamine has been proven to regulate axon regeneration in C. elegans, but its role in mammals is still unknown.
OBJECTIVE: To explore the effect of N-arachidonylethanolamine on axon regeneration of mouse dorsal root ganglion neurons.
METHODS: Dorsal root ganglion tissues were taken from L4-L5 of ICR mice aged 6-8 weeks, digested with collagenase and trypsin, and divided into URB597 group, N-arachidonylethanolamine group, methyl-sulfoxide control group, and electroporated green fluorescent protein and specific siRNA mixture group. After 3 days of in vitro culture, the axons were labeled by neuron-specific immunoassay. Axon length, and the number of primary and secondary branches were measured, and the statistics was performed to determine the regulatory effect of N-arachidonylethanolamine on the regeneration of dorsal root ganglion neuron axon.
RESULTS AND CONCLUSION: (1) After inhibiting fatty acid amide hydrolase activity, there was no obvious change in dorsal root ganglion cell axon regeneration and URB597 had no cytotoxic effect on dorsal root ganglion neuron cells (P > 0.05). (2) After fatty acid amide hydrolase knockdown, dorsal root ganglion cell axon regeneration had no obvious change (P > 0.05). (3) Exogenous N-arachidonylethanolamine had no obvious regulatory effect on dorsal root ganglion cell axon regeneration and N-arachidonylethanolamine had no cytotoxic effect on dorsal root ganglion neuron cells (P > 0.05). (4) Inhibiting fatty acid amide hydrolase activity or adding exogenous N-arachidonylethanolamine has no effect on dorsal root ganglion neuron axon branches (P > 0.05).

Key words: dorsal root ganglion, neurons, N-arachidylethanolamine, fatty acid amide hydrolase, axon regeneration, cytotoxicity

CLC Number:

Zhang Haonan, Wang Xingran, Li Meimei, Ma Jinjin, Ma Yanxia, Saijilafu. Effect of N-arachidonylethanolamine on axon regeneration of the dorsal root ganglion[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(25): 3999-4003.

Figures/Tables 4

References

[1] ZOU S, KUMAR U. Cannabinoid Receptors and the Endocannabinoid System: Signaling and Function in the Central Nervous System. Int J Mol Sci. 2018;19(3):833.
[2] KRISHNAN G, CHATTERJEE N. Differential immune mechanism to HIV-1 Tat variants and its regulation by AEA [corrected]. Sci Rep. 2015;5:9887.
[3] ZAJKOWSKA Z, BORSINI A, NIKKHESLAT N, et al. Differential effect of interferon-alpha treatment on AEA and 2-AG levels. Brain Behav Immun. 2020;90:248-258.
[4] MACCARRONE M, ROSSI S, BARI M, et al. Anandamide inhibits metabolism and physiological actions of 2-arachidonoylglycerol in the striatum. Nat Neurosci. 2008; 11(2):152-159.
[5] LITTLE TJ, CVIJANOVIC N, DIPATRIZIO NV, et al. Plasma endocannabinoid levels in lean, overweight, and obese humans: relationships to intestinal permeability markers, inflammation, and incretin secretion. Am J Physiol Endocrinol Metab. 2018; 315(4):E489-E495.
[6] LI H, YIN J, ZHANG Z, et al. Auricular vagal nerve stimulation ameliorates burn-induced gastric dysmotility via sympathetic-COX-2 pathways in rats. Neurogastroenterol Motil. 2016;28(1):36-42.
[7] MALEK N, KUCHARCZYK M, STAROWICZ K. Alterations in the anandamide metabolism in the development of neuropathic pain. Biomed Res Int. 2014;2014:686908.
[8] KAMIMURA R, HOSSAIN MZ, UNNO S, et al. Inhibition of 2-arachydonoylgycerol degradation attenuates orofacial neuropathic pain in trigeminal nerve-injured mice. J Oral Sci. 2018;60(1):37-44.
[9] OAKES MD, LAW WJ, CLARK T, et al. Cannabinoids Activate Monoaminergic Signaling to Modulate Key C. elegans Behaviors. J Neurosci. 2017;37(11):2859-2869.
[10] FELIÚ A, BONILLA DEL RÍO I, CARRILLO-SALINAS FJ, et al. 2-Arachidonoylglycerol Reduces Proteoglycans and Enhances Remyelination in a Progressive Model of Demyelination. J Neurosci. 2017;37(35):8385-8398.
[11] MECHA M, YANGUAS-CASÁS N, FELIÚ A, et al. The endocannabinoid 2-AG enhances spontaneous remyelination by targeting microglia. Brain Behav Immun. 2019;77:110-126.
[12] TAPIA M, DOMINGUEZ A, ZHANG W, et al. Cannabinoid Receptors Modulate Neuronal Morphology and AnkyrinG Density at the Axon Initial Segment. Front Cell Neurosci. 2017;11:5.
[13] WOOLUMS BM, MCCRAY BA, SUNG H, et al. TRPV4 disrupts mitochondrial transport and causes axonal degeneration via a CaMKII-dependent elevation of intracellular Ca2. Nat Commun. 2020;11(1):2679.
[14] HE JC, GOMES I, NGUYEN T, et al. The G alpha(o/i)-coupled cannabinoid receptor-mediated neurite outgrowth involves Rap regulation of Src and Stat3. J Biol Chem. 2005;280(39):33426-33434.
[15] JANG Y, JUNG J, KIM H, et al. Axonal neuropathy-associated TRPV4 regulates neurotrophic factor-derived axonal growth. J Biol Chem. 2012;287(8):6014-6024.
[16] GUGLIANDOLO E, D’AMICO R, CORDARO M, et al. Effect of PEA-OXA on neuropathic pain and functional recovery after sciatic nerve crush. J Neuroinflammation. 2018;15(1):264.
[17] PASTUHOV SI, FUJIKI K, NIX P, et al. Endocannabinoid-Goα signalling inhibits axon regeneration in Caenorhabditis elegans by antagonizing Gqα-PKC-JNK signalling. Nat Commun. 2012;3:1136.
[18] FORNARO M, SHARTHIYA H, TIWARI V. Adult Mouse DRG Explant and Dissociated Cell Models to Investigate Neuroplasticity and Responses to Environmental Insults Including Viral Infection. J Vis Exp. 2018;(133):56757.
[19] LIN YT, CHEN JC. Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release. J Vis Exp. 2018;(140):57569.
[20] MALIN SA, DAVIS BM, MOLLIVER DC. Production of dissociated sensory neuron cultures and considerations for their use in studying neuronal function and plasticity. Nat Protoc. 2007;2(1):152-160.
[21] DAVIES A, LUMSDEN A. Relation of target encounter and neuronal death to nerve growth factor responsiveness in the developing mouse trigeminal ganglion. J Comp Neurol. 1984;223(1):124-137.
[22] LINDWALL C, KANJE M. The Janus role of c-Jun: cell death versus survival and regeneration of neonatal sympathetic and sensory neurons. Exp Neurol. 2005;196(1):184-194.
[23] MA JJ, JU X, XU RJ, et al. Telomerase Reverse Transcriptase and p53 Regulate Mammalian Peripheral Nervous System and CNS Axon Regeneration Downstream of c-Myc. J Neurosci. 2019;39(46):9107-9118.
[24] SAIJILAFU, ZHANG BY, ZHOU FQ. In vivo electroporation of adult mouse sensory neurons for studying peripheral axon regeneration. Methods Mol Biol. 2014;1162:167-175.
[25] ZHANG BY, SAIJILAFU, LIU CM, et al. Akt-independent GSK3 inactivation downstream of PI3K signaling regulates mammalian axon regeneration. Biochem Biophys Res Commun. 2014;443(2):743-748.
[26] COSTA FA, MOREIRA NETO FL. Satellite glial cells in sensory ganglia: its role in pain. Rev Bras Anestesiol. 2015;65(1):73-81.
[27] AVRAHAM O, DENG PY, JONES S, et al. Satellite glial cells promote regenerative growth in sensory neurons. Nat Commun. 2020;11(1):4891.
[28] BAI J, LIU F, WU LF, et al. Attenuation of TRPV1 by AMG-517 after nerve injury promotes peripheral axonal regeneration in rats. Mol Pain. 2018;14:1744806918777614.
[29] WINKLER K, RAMER R, DITHMER S, et al. Fatty acid amide hydrolase inhibitors confer anti-invasive and antimetastatic effects on lung cancer cells. Oncotarget. 2016;7(12):15047-15064.
[30] LEVER IJ, ROBINSON M, CIBELLI M, et al. Localization of the endocannabinoid-degrading enzyme fatty acid amide hydrolase in rat dorsal root ganglion cells and its regulation after peripheral nerve injury. J Neurosci. 2009;29(12):3766-3780.
[31] SAIJILAFU, HUR EM, LIU CM, et al. PI3K-GSK3 signalling regulates mammalian axon regeneration by inducing the expression of Smad1. Nat Commun. 2013;4:2690.
[32] HE XL, YANG L, WANG ZJ, et al. Solid lipid nanoparticles loading with curcumin and dexanabinol to treat major depressive disorder. Neural Regen Res. 2021;16(3):537-542.
[33] PACHER P, BÁTKAI S, KUNOS G. The endocannabinoid system as an emerging target of pharmacotherapy. Pharmacol Rev. 2006;58(3):389-462.