2ef80bf3fc2654629b916b6e11caa113  pmc_35159165.pdf
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02d8caa25c328c40e4f8f6f907709b2ff7680de1a355270b55dae4b1697c7d3d  pmc_35159165.pdf

Title:           Neuron–Microglia Contact-Dependent Mechanisms Attenuate Methamphetamine-Induced Microglia Reactivity and Enhance Neuronal Plasticity
Subject:         Exposure to methamphetamine (Meth) has been classically associated with damage to neuronal terminals. However, it is now becoming clear that addiction may also result from the interplay between glial cells and neurons. Recently, we demonstrated that binge Meth administration promotes microgliosis and microglia pro-inflammation via astrocytic glutamate release in a TNF/IP3R2-Ca2+-dependent manner. Here, we investigated the contribution of neuronal cells to this process. As the crosstalk between microglia and neurons may occur by contact-dependent and/or contact-independent mechanisms, we developed co-cultures of primary neurons and microglia in microfluidic devices to investigate how their interaction affects Meth-induced microglia activation. Our results show that neurons exposed to Meth do not activate microglia in a cell-autonomous way but require astrocyte mediation. Importantly, we found that neurons can partially prevent Meth-induced microglia activation via astrocytes, which seems to be achieved by increasing arginase 1 expression and strengthening the CD200/CD200r pathway. We also observed an increase in synaptic individual area, as determined by co-localization of pre- and post-synaptic markers. The present study provides evidence that contact-dependent mechanisms between neurons and microglia can attenuate pro-inflammatory events such as Meth-induced microglia activation.
Keywords:        methamphetamine; neuron-to-microglia; neuroprotection; contact-dependent; CD200; PSD95
Author:          Joana Bravo, Inês Ribeiro, Ana Filipa Terceiro, Elva B. Andrade, Camila Cabral Portugal, Igor M. Lopes, Maria M. Azevedo, Mafalda Sousa, Cátia D. F. Lopes, Andrea C. Lobo, Teresa Canedo, João Bettencourt Relvas and Teresa Summavielle
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      Title: Introduction 
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     Item: 
      Title: Materials and Methods 
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      Children: 
       Item: 
        Title: Animals 
        Destination: subsection.2.1
       Item: 
        Title: Neuronal Cell Culture 
        Destination: subsection.2.2
       Item: 
        Title: Microglia and Astrocyte Purified Cultures Obtained from Mixed Glial Cell Cultures 
        Destination: subsection.2.3
       Item: 
        Title: Microglia Incubation with Neuron-Conditioned Medium 
        Destination: subsection.2.4
       Item: 
        Title: Co-Cultures of Neurons and Microglia in Microfluidic Devices 
        Destination: subsection.2.5
       Item: 
        Title: Astrocytic-Conditioned Medium 
        Destination: subsection.2.6
       Item: 
        Title: Treatment of Neuron Microglia Co-Cultures 
        Destination: subsection.2.7
       Item: 
        Title: Immunocytochemistry and Image Acquisition 
        Destination: subsection.2.8
       Item: 
        Title: Phagocytic Assay 
        Destination: subsection.2.9
       Item: 
        Title: Fluorescence Intensity Quantification 
        Destination: subsection.2.10
       Item: 
        Title: RNA Extraction, cDNA Synthesis, and qRT-PCR 
        Destination: subsection.2.11
       Item: 
        Title: Flow Cytometry of Microglia 
        Destination: subsection.2.12
       Item: 
        Title: Quantification of Synaptic Proteins 
        Destination: subsection.2.13
       Item: 
        Title: Statistical Analysis 
        Destination: subsection.2.14
     Item: 
      Title: Results 
      Destination: section.3
      Children: 
       Item: 
        Title: Neurons Exposed to Meth Do Not Promote Microglial Activation through Contact-Dependent or -Independent Mechanisms 
        Destination: subsection.3.1
       Item: 
        Title: Microglia Activation Induced by Meth-Exposed Astrocytes Is Partially Prevented by Neurons 
        Destination: subsection.3.2
       Item: 
        Title: Neurons Increased Self-Protection from Meth-Induced Activation of Microglia through Contact-Dependent Mechanisms 
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       Item: 
        Title: Synaptic Proteins Expression Increases in Co-Cultures of Neurons and ACM Meth-Microglia 
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     Item: 
      Title: Discussion 
      Destination: section.4
     Item: 
      Title: Conclusions 
      Destination: section.5
     Item: 
      Title: References
      Destination: section.5
   Info: 
    Title: Neuron Microglia Contact-Dependent Mechanisms Attenuate Methamphetamine-Induced Microglia Reactivity and Enhance Neuronal Plasticity
    Author: Joana Bravo, Inês Ribeiro, Ana Filipa Terceiro, Elva B. Andrade, Camila Cabral Portugal, Igor M. Lopes, Maria M. Azevedo, Mafalda Sousa, Cátia D. F. Lopes, Andrea C. Lobo, Teresa Canedo, João Bettencourt Relvas and Teresa Summavielle
    Subject: Exposure to methamphetamine (Meth) has been classically associated with damage to neuronal terminals. However, it is now becoming clear that addiction may also result from the interplay between glial cells and neurons. Recently, we demonstrated that binge Meth administration promotes microgliosis and microglia pro-inflammation via astrocytic glutamate release in a TNF/IP3R2-Ca2+-dependent manner. Here, we investigated the contribution of neuronal cells to this process. As the crosstalk between microglia and neurons may occur by contact-dependent and/or contact-independent mechanisms, we developed co-cultures of primary neurons and microglia in microfluidic devices to investigate how their interaction affects Meth-induced microglia activation. Our results show that neurons exposed to Meth do not activate microglia in a cell-autonomous way but require astrocyte mediation. Importantly, we found that neurons can partially prevent Meth-induced microglia activation via astrocytes, which seems to be achieved by increasing arginase 1 expression and strengthening the CD200/CD200r pathway. We also observed an increase in synaptic individual area, as determined by co-localization of pre- and post-synaptic markers. The present study provides evidence that contact-dependent mechanisms between neurons and microglia can attenuate pro-inflammatory events such as Meth-induced microglia activation.
    Keywords: methamphetamine; neuron-to-microglia; neuroprotection; contact-dependent; CD200; PSD95
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  Checksum: 2908f871
   Type: CRC32
  Checksum: 2ef80bf3fc2654629b916b6e11caa113
   Type: MD5
  Checksum: 91b1e9305cf733dc5e6ba8d015d2b935e02775b4
   Type: SHA-1
  Checksum: 02d8caa25c328c40e4f8f6f907709b2ff7680de1a355270b55dae4b1697c7d3d
   Type: SHA-256