Targeting CB1 and GPR55 Endocannabinoid Receptors as a Potential Neuroprotective Approach for Parkinson’s Disease

Abstract

Cannabinoid CB1 receptors (CB1R) and the GPR55 receptor are expressed in striatum and are potential targets in the therapy of Parkinson’s disease (PD), one of the most prevalent neurodegenerative diseases in developed countries. The aim of this paper was to address the potential of ligands acting on those receptors to prevent the action of a neurotoxic agent, MPP+, that specifically affects neurons of the substantia nigra due to uptake via the dopamine DAT transporter. The SH-SY5Y cell line model was used as it expresses DAT and, therefore, is able to uptake MPP+ that inhibits complex I of the respiratory mitochondrial chain and leads to cell death. Cells were transfected with cDNAs coding for either or both receptors. Receptors in cotransfected cells formed heteromers as indicated by the in situ proximity ligation assays. Cell viability was assayed by oxygen rate consumption and by the bromide-based MTT method. Assays of neuroprotection using two concentrations of MPP+ showed that cells expressing receptor heteromers were more resistant to the toxic effect. After correction by effects on cell proliferation, the CB1R antagonist, SR141716, afforded an almost full neuroprotection in CB1R-expressing cells even when a selective agonist, ACEA, was present. In contrast, SR141716 was not effective in cells expressing CB1/GPR55 heteromeric complexes. In addition, an agonist of GPR55, CID1792197, did not enhance neuroprotection in GPR55-expressing cells. These results show that neurons expressing heteromers are more resistant to cell death but question the real usefulness of CB1R, GPR55, and their heteromers as targets to afford PD-related neuroprotection.

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Funding

This study was funded by Fundació La Marató de TV3 (Grant Numbers 20141330 and 20141331).

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Correspondence to Eva Martínez-Pinilla or José Luis Lanciego or Rafael Franco.

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Supplementary Fig. 1
figure9

MTT reduction assay in SH-SY5Y (a), SH-SY5Y-CB1R (b) and SH-SY5Y-GPR55 (c) cells treated with MPP+ (1-2 mM) for 48 h. Cell damage is represented as the percentage of MTT reduction versus control. Data are the mean ± SEM of five independent experiments. Significant differences were analyzed by a one-way ANOVA followed by post-hoc Tukey’s test. ***p < 0.001 compared with control. (PNG 332 kb)

Supplementary Fig. 2
figure10

MTT reduction assay in SH-SY5Y (a) and SH-SY5Y-CB1R/GPR55 (b) cells treated with MPP+ (1-2 mM) for 48 h. Cell damage is represented as the percentage of MTT reduction versus control. Data are the mean ± SEM of five independent experiments. Significant differences were analyzed by a one-way ANOVA followed by post-hoc Tukey’s test. ***p < 0.001 compared with control. (PNG 243 kb)

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Martínez-Pinilla, E., Aguinaga, D., Navarro, G. et al. Targeting CB1 and GPR55 Endocannabinoid Receptors as a Potential Neuroprotective Approach for Parkinson’s Disease. Mol Neurobiol 56, 5900–5910 (2019). https://doi.org/10.1007/s12035-019-1495-4

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Keywords

  • Cannabinoid receptors
  • GPCRs
  • Neurodegenerative diseases
  • Neuroprotection
  • MPP+