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Repeated cocaine administration upregulates CB₂ receptor expression in striatal medium-spiny neurons that express dopamine D₁ receptors in mice.

ABSTRACT: Cannabinoid CB₂ receptors (CB₂R) are importantly involved in drug reward and addiction. However, the cellular mechanisms underlying CB₂R action remain unclear. We have previously reported that cocaine self-administration upregulates CB₂R expression in midbrain dopamine (DA) neurons. In the present study, we investigated whether cocaine or heroin also alters CB₂R expression in striatal medium-spiny neurons that express dopamine D₁ or D₂ receptors (D₁-MSNs, D₂-MSNs) and microglia. Due to the concern of CB₂R antibody specificity, we developed three mouse CB₂-specific probes to detect CB₂R mRNA using quantitative RT-PCR and RNAscope in situ hybridization (ISH) assays. We found that a single injection of cocaine failed to alter, while repeated cocaine injections or self-administration dose-dependently upregulated CB₂R gene expression in both brain (cortex and striatum) and periphery (spleen). In contrast, repeated administration of heroin produced a dose-dependent reduction in striatal CB₂ mRNA expression. RNAscope ISH assays detected CB₂R mRNA in striatal D₁- and D₂-MSNs, not in microglia. We then used transgenic CX3CR1^eGFP/+ microglia reporter mice and D₁- or D₂-Cre-RiboTag mice to purify striatal microglia or ribosome-associated mRNAs from CX3CR1^eGFP/+, D₁-MSNs, or D₂-MSNs, respectively. We found that CB₂R upregulation occurred mainly in D₁-MSNs, not in D₂-MSNs or microglia, in the nucleus accumbens rather than the dorsal striatum. These findings indicate that repeated cocaine exposure may upregulate CB₂R expression in both brain and spleen, with regional and cell type-specific profiles. In the striatum, CB₂R upregulation occurs mainly in D₁-MSNs in the nucleus accumbens. Given the important role of D₁-MSNs in brain reward function, the present findings provide new insight into mechanisms by which brain CB₂Rs modulate cocaine action.

SUBMITTER: Zhang HY

PROVIDER: S-EPMC8975868 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature

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Repeated cocaine administration upregulates CB2 receptor expression in striatal medium-spiny neurons that express dopamine D1 receptors in mice.

Zhang Hai-Ying HY De Biase Lindsay L Chandra Ramesh R Shen Hui H Liu Qing-Rong QR Gardner Eliot E Lobo Mary Kay MK Xi Zheng-Xiong ZX

Acta pharmacologica Sinica 20210727 4

Cannabinoid CB2 receptors (CB2R) are importantly involved in drug reward and addiction. However, the cellular mechanisms underlying CB2R action remain unclear. We have previously reported that cocaine self-administration upregulates CB2R expression in midbrain dopamine (DA) neurons. In the present study, we investigated whether cocaine or heroin also alters CB2R expression in striatal medium-spiny neurons that express dopamine D1 ...[more]

PMID: 34316031

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Project description:The classical motor symptoms of Parkinson's disease (PD) are caused by degeneration of dopaminergic neurons in the substantia nigra, which is followed by secondary dendritic pruning and spine loss at striatal medium spiny neurons (MSN). We hypothesize that these morphological changes at MSN underlie at least in part long-term motor complications in PD patients. In order to define the potential benefits and limitations of dopamine substitution, we tested in a mouse model whether dendritic pruning and spine loss can be reversible when dopaminergic axon terminals regenerate. In order to induce degeneration of nigrostriatal dopaminergic neurons we used the toxicity of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in C57BL/6J mice; 30 mg/kg MPTP was applied i.p. on five consecutive days. In order to assess the consequences of dopamine depletion, mice were analyzed 21 days after the last injection. In order to test reversibility of MSN changes we exploited the property of this model that striatal axon terminals regenerate by sprouting within 90 days and analyzed a second cohort 90 days after MPTP. Degeneration of dopaminergic neurons was confirmed by counting TH-positive neurons in the substantia nigra and by analyzing striatal catecholamines. Striatal catecholamine recovered 90 days after MPTP. MSN morphology was visualized by Golgi staining and quantified as total dendritic length, number of dendritic branch points, and density of dendritic spines. All morphological parameters of striatal MSN were reduced 21 days after MPTP. Statistical analysis indicated that dendritic pruning and the reduction of spine density represent two distinct responses to dopamine depletion. Ninety days after MPTP, all morphological changes recovered. Our findings demonstrate that morphological changes in striatal MSN resulting from dopamine depletion are reversible. They suggest that under optimal conditions, symptomatic dopaminergic therapy might be able to prevent maladaptive plasticity and long-term motor complications in PD patients.

Project description:Many neurological disorders and diseases including drug addiction are associated with specific neuronal cell types in the brain. The striatum, a region that plays a critically important role in the development of addictive drug-related behavior, provides a good example of the cellular heterogeneity challenges associated with analyses of specific neuronal cell types. Such studies are needed to identify the adaptive changes in neuroproteomic signaling that occur in response to diseases such as addiction. The striatum contains two major cell types, D1 and D2 type dopaminoceptive medium spiny neurons (MSNs), whose cell bodies and processes are intermingled throughout this region. Since little is known about the proteomes of these two neuronal cell populations, we have begun to address this challenge by using fluorescence-activated nuclear sorting (FANS) to isolate nuclei-containing fractions from striatum from D1 and D2 "Translating Ribosome Affinity Purification" (TRAP) mice. This approach enabled us to devise and implement a robust and reproducible workflow for preparing samples from specific MSN cell types for mass spectrometry analyses. These analyses quantified at least 685 proteins in each of four biological replicates of 50 K sorted nuclei from two D1 mice/replicate and from each of four biological replicates of 50 K sorted nuclei from two D2 mice/replicate. Proteome analyses identified 87 proteins that were differentially expressed in D1 versus D2 MSN nuclei and principal component analysis (PCA) of these proteins separated the 8 biological replicates into specific cell types. Central network analysis of the 87 differentially expressed proteins identified Hnrnpd and Hnmpa2b1 in D1 and Cct2 and Cct7 in D2 as potential central interactors. This workflow can now be used to improve our understanding of many neurological diseases including characterizing the short and long-term impact of drugs of abuse on the proteomes of these two dopaminoceptive neuronal populations.

Dataset Information

Repeated cocaine administration upregulates CB₂ receptor expression in striatal medium-spiny neurons that express dopamine D₁ receptors in mice.

Publications

Repeated cocaine administration upregulates CB<sub>2</sub> receptor expression in striatal medium-spiny neurons that express dopamine D<sub>1</sub> receptors in mice.

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