Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells
Regeneration of peripheral nerves depends on the ability of axons to navigate through an altered extracellular environment. It has been suggested that Schwann cells facilitate this process through their secretion of neuropeptides and proteases. Using the RT4-D6P2T Schwann cell line (RT4), we have pr...
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MDPI AG
2021-08-01
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| author | Sarah Thomas Broome Giuseppe Musumeci Alessandro Castorina |
| author_facet | Sarah Thomas Broome Giuseppe Musumeci Alessandro Castorina |
| author_sort | Sarah Thomas Broome |
| collection | DOAJ |
| description | Regeneration of peripheral nerves depends on the ability of axons to navigate through an altered extracellular environment. It has been suggested that Schwann cells facilitate this process through their secretion of neuropeptides and proteases. Using the RT4-D6P2T Schwann cell line (RT4), we have previously shown that RT4 cultures endogenously express the neuropeptide PACAP, and respond to exogenous stimulation by inducing the expression of tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA) via PAC1 receptor activation. In this study, based on recent findings showing that doxycycline and minocycline act as positive allosteric modulators (PAMs) of the PAC1 receptor, we tested if treatment with these tetracyclines induced the expression and activity of tPA and uPA in RT4 cells. Using ELISA and zymographic analyses, we demonstrate that doxycycline and minocycline reliably induce the secretion and activity of both tPA and uPA, which is paralleled by increased expression levels, as shown by immunocytochemistry and Western blots. These actions were mediated, at least in part, by the PAC1 receptor, as PACAP6-38 mitigated tetracycline-induced expression and activity of tPA and uPA. We conclude that doxycycline and minocycline act as PAMs of the PAC1 receptor to promote proteolytic activity in RT4 cells. |
| first_indexed | 2024-03-10T09:01:46Z |
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| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-10T09:01:46Z |
| publishDate | 2021-08-01 |
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| spelling | doaj.art-d2241311b67b407cafa83ec1dd5cbfc12023-11-22T06:45:11ZengMDPI AGApplied Sciences2076-34172021-08-011116767310.3390/app11167673Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann CellsSarah Thomas Broome0Giuseppe Musumeci1Alessandro Castorina2Laboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaDepartment of Biomedical and Biotechnological Sciences, Anatomy, Histology and Movement Sciences Section, School of Medicine, University of Catania, 95125 Catania, ItalyLaboratory of Cellular and Molecular Neuroscience (LCMN), School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, AustraliaRegeneration of peripheral nerves depends on the ability of axons to navigate through an altered extracellular environment. It has been suggested that Schwann cells facilitate this process through their secretion of neuropeptides and proteases. Using the RT4-D6P2T Schwann cell line (RT4), we have previously shown that RT4 cultures endogenously express the neuropeptide PACAP, and respond to exogenous stimulation by inducing the expression of tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA) via PAC1 receptor activation. In this study, based on recent findings showing that doxycycline and minocycline act as positive allosteric modulators (PAMs) of the PAC1 receptor, we tested if treatment with these tetracyclines induced the expression and activity of tPA and uPA in RT4 cells. Using ELISA and zymographic analyses, we demonstrate that doxycycline and minocycline reliably induce the secretion and activity of both tPA and uPA, which is paralleled by increased expression levels, as shown by immunocytochemistry and Western blots. These actions were mediated, at least in part, by the PAC1 receptor, as PACAP6-38 mitigated tetracycline-induced expression and activity of tPA and uPA. We conclude that doxycycline and minocycline act as PAMs of the PAC1 receptor to promote proteolytic activity in RT4 cells.https://www.mdpi.com/2076-3417/11/16/7673pituitary adenylate cyclase-activating peptide (PACAP)PAC1 receptordoxycyclineminocyclinepositive allosteric modulatorSchwann cells |
| spellingShingle | Sarah Thomas Broome Giuseppe Musumeci Alessandro Castorina Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells Applied Sciences pituitary adenylate cyclase-activating peptide (PACAP) PAC1 receptor doxycycline minocycline positive allosteric modulator Schwann cells |
| title | Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells |
| title_full | Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells |
| title_fullStr | Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells |
| title_full_unstemmed | Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells |
| title_short | Doxycycline and Minocycline Act as Positive Allosteric Modulators of the PAC1 Receptor and Induce Plasminogen Activators in RT4 Schwann Cells |
| title_sort | doxycycline and minocycline act as positive allosteric modulators of the pac1 receptor and induce plasminogen activators in rt4 schwann cells |
| topic | pituitary adenylate cyclase-activating peptide (PACAP) PAC1 receptor doxycycline minocycline positive allosteric modulator Schwann cells |
| url | https://www.mdpi.com/2076-3417/11/16/7673 |
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