Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells
Buspirone is an anxiolytic drug with robust serotonin receptor 1A (Htr1a) agonist activities. However, evidence has demonstrated that this drug also targets the dopamine D3 receptor (Drd3), where it acts as a potent antagonist. In vivo, Drd3 blockade is neuroprotective and reduces inflammation in mo...
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MDPI AG
2021-05-01
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| author | Sarah Thomas Broome Teagan Fisher Alen Faiz Kevin A. Keay Giuseppe Musumeci Ghaith Al-Badri Alessandro Castorina |
| author_facet | Sarah Thomas Broome Teagan Fisher Alen Faiz Kevin A. Keay Giuseppe Musumeci Ghaith Al-Badri Alessandro Castorina |
| author_sort | Sarah Thomas Broome |
| collection | DOAJ |
| description | Buspirone is an anxiolytic drug with robust serotonin receptor 1A (Htr1a) agonist activities. However, evidence has demonstrated that this drug also targets the dopamine D3 receptor (Drd3), where it acts as a potent antagonist. In vivo, Drd3 blockade is neuroprotective and reduces inflammation in models of Parkinson’s disease. To test if buspirone also elicited anti-inflammatory activities in vitro, we generated stable Drd3<sup>−/−</sup> and Htr1a<sup>−/−</sup> BV2 microglial cell lines using CRISPR-Cas9 technology and then tested the effects of buspirone after lipopolysaccharide (LPS) challenge. We found that LPS exposure had no effect on cell viability, except in Htr1a<sup>−/−</sup> cells, where viability was reduced (<i>p</i> < 0.001). Drug treatment reduced viability in Drd3<sup>−/−</sup> cells, but not in WT or Htr1a<sup>−/−</sup> cells. Buspirone counteracted LPS-induced NO release, NOS2, IL-1β and TNF-α gene expression in WT cells, whereas it exerted limited effects in Drd3<sup>−/−</sup> or Htr1a<sup>−/−</sup> microglia. In summary, our findings indicate that buspirone attenuates microglial polarization after LPS challenge. These results also highlight some major effects of Drd3 or Htr1a genetic ablation on microglial biology, raising important questions on the complex role of neurotransmitters in regulating microglia functions. |
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| issn | 2073-4409 |
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| last_indexed | 2024-03-10T11:04:43Z |
| publishDate | 2021-05-01 |
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| series | Cells |
| spelling | doaj.art-b76bb09ef3cf477aaa8128b6e3c7f03c2023-11-21T21:16:50ZengMDPI AGCells2073-44092021-05-01106131210.3390/cells10061312Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial CellsSarah Thomas Broome0Teagan Fisher1Alen Faiz2Kevin A. Keay3Giuseppe Musumeci4Ghaith Al-Badri5Alessandro Castorina6Laboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaLaboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaRespiratory Bioinformatics and Molecular Biology (RBMB) Group, School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaLaboratory of Neural Structure and Function (LNSF), School of Medical Sciences (Neuroscience), Faculty of Medicine and Health, University of Sydney, Sydney, NSW 2006, AustraliaSection of Human Anatomy and Histology, Department of Biomedical and Biotechnological Sciences, University of Catania, via S. Sofia, 87, 95123 Catania, ItalyLaboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaLaboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaBuspirone is an anxiolytic drug with robust serotonin receptor 1A (Htr1a) agonist activities. However, evidence has demonstrated that this drug also targets the dopamine D3 receptor (Drd3), where it acts as a potent antagonist. In vivo, Drd3 blockade is neuroprotective and reduces inflammation in models of Parkinson’s disease. To test if buspirone also elicited anti-inflammatory activities in vitro, we generated stable Drd3<sup>−/−</sup> and Htr1a<sup>−/−</sup> BV2 microglial cell lines using CRISPR-Cas9 technology and then tested the effects of buspirone after lipopolysaccharide (LPS) challenge. We found that LPS exposure had no effect on cell viability, except in Htr1a<sup>−/−</sup> cells, where viability was reduced (<i>p</i> < 0.001). Drug treatment reduced viability in Drd3<sup>−/−</sup> cells, but not in WT or Htr1a<sup>−/−</sup> cells. Buspirone counteracted LPS-induced NO release, NOS2, IL-1β and TNF-α gene expression in WT cells, whereas it exerted limited effects in Drd3<sup>−/−</sup> or Htr1a<sup>−/−</sup> microglia. In summary, our findings indicate that buspirone attenuates microglial polarization after LPS challenge. These results also highlight some major effects of Drd3 or Htr1a genetic ablation on microglial biology, raising important questions on the complex role of neurotransmitters in regulating microglia functions.https://www.mdpi.com/2073-4409/10/6/1312microgliadopamine D3 receptor5-hydroxytryptamine 1a receptorneuroinflammationParkinson’s disease |
| spellingShingle | Sarah Thomas Broome Teagan Fisher Alen Faiz Kevin A. Keay Giuseppe Musumeci Ghaith Al-Badri Alessandro Castorina Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells Cells microglia dopamine D3 receptor 5-hydroxytryptamine 1a receptor neuroinflammation Parkinson’s disease |
| title | Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells |
| title_full | Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells |
| title_fullStr | Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells |
| title_full_unstemmed | Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells |
| title_short | Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells |
| title_sort | assessing the anti inflammatory activity of the anxiolytic drug buspirone using crispr cas9 gene editing in lps stimulated bv 2 microglial cells |
| topic | microglia dopamine D3 receptor 5-hydroxytryptamine 1a receptor neuroinflammation Parkinson’s disease |
| url | https://www.mdpi.com/2073-4409/10/6/1312 |
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