Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling
Background/Aims: The neutral, non-essential amino acid glycine has manifold functions and effects under physiological and pathophysiological conditions. Besides its function as a neurotransmitter in the central nervous system, glycine also exerts immunomodulatory effects and as an osmolyte it partic...
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| Format: | Article |
| Language: | English |
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Cell Physiol Biochem Press GmbH & Co KG
2018-10-01
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| Series: | Cellular Physiology and Biochemistry |
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| Online Access: | https://www.karger.com/Article/FullText/494646 |
| _version_ | 1819118671536062464 |
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| author | Michael Kittl Heidemarie Dobias Marlena Beyreis Tobias Kiesslich Christian Mayr Martin Gaisberger Markus Ritter Hubert H. Kerschbaum Martin Jakab |
| author_facet | Michael Kittl Heidemarie Dobias Marlena Beyreis Tobias Kiesslich Christian Mayr Martin Gaisberger Markus Ritter Hubert H. Kerschbaum Martin Jakab |
| author_sort | Michael Kittl |
| collection | DOAJ |
| description | Background/Aims: The neutral, non-essential amino acid glycine has manifold functions and effects under physiological and pathophysiological conditions. Besides its function as a neurotransmitter in the central nervous system, glycine also exerts immunomodulatory effects and as an osmolyte it participates in cell volume regulation. During phagocytosis, glycine contributes to (local) cell volume-dependent processes like lamellipodium formation. Similar to the expansion of the lamellipodium we assume that glycine also affects the migration of microglial cells in a cell volume-dependent manner. Methods: Mean cell volume (MCV) and cell migration were determined using flow cytometry and trans-well migration assays, respectively. Electrophysiological recordings of the cell membrane potential (Vmem) and swelling-dependent chloride (Cl-) currents (IClswell, VSOR, VRAC) were performed using the whole-cell patch clamp technique. Results: In the murine microglial cell line BV-2, flow cytometry analysis revealed that glycine (5 mM) increases the MCV by ∼9%. The glycine-dependent increase in MCV was suppressed by the partial sodium-dependent neutral amino acid transporter (SNAT) antagonist MeAIB and augmented by the Cl- current blocker DCPIB. Electrophysiological recordings showed that addition of glycine activates a Cl- current under isotonic conditions resembling features of the swelling-activated Cl- current (IClswell). The cell membrane potential (Vmem) displayed a distinctive time course after glycine application; initially, glycine evoked a rapid depolarization mediated by Na+-coupled glycine uptake via SNAT, followed by a further gradual depolarization, which was fully suppressed by DCPIB. Interestingly, glycine significantly increased migration of BV-2 cells, which was suppressed by MeAIB, suggesting that SNAT is involved in the migration process of microglial cells. Conclusion: We conclude that glycine acts as a chemoattractant for microglial cells presumably by a cell volume-dependent mechanism involving SNAT-mediated cell swelling. |
| first_indexed | 2024-12-22T05:52:35Z |
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| id | doaj.art-3f68b6d4a51b41f3ad84825f5d1d85bc |
| institution | Directory Open Access Journal |
| issn | 1015-8987 1421-9778 |
| language | English |
| last_indexed | 2024-12-22T05:52:35Z |
| publishDate | 2018-10-01 |
| publisher | Cell Physiol Biochem Press GmbH & Co KG |
| record_format | Article |
| series | Cellular Physiology and Biochemistry |
| spelling | doaj.art-3f68b6d4a51b41f3ad84825f5d1d85bc2022-12-21T18:36:50ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-10-015041460147310.1159/000494646494646Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell SwellingMichael KittlHeidemarie DobiasMarlena BeyreisTobias KiesslichChristian MayrMartin GaisbergerMarkus RitterHubert H. KerschbaumMartin JakabBackground/Aims: The neutral, non-essential amino acid glycine has manifold functions and effects under physiological and pathophysiological conditions. Besides its function as a neurotransmitter in the central nervous system, glycine also exerts immunomodulatory effects and as an osmolyte it participates in cell volume regulation. During phagocytosis, glycine contributes to (local) cell volume-dependent processes like lamellipodium formation. Similar to the expansion of the lamellipodium we assume that glycine also affects the migration of microglial cells in a cell volume-dependent manner. Methods: Mean cell volume (MCV) and cell migration were determined using flow cytometry and trans-well migration assays, respectively. Electrophysiological recordings of the cell membrane potential (Vmem) and swelling-dependent chloride (Cl-) currents (IClswell, VSOR, VRAC) were performed using the whole-cell patch clamp technique. Results: In the murine microglial cell line BV-2, flow cytometry analysis revealed that glycine (5 mM) increases the MCV by ∼9%. The glycine-dependent increase in MCV was suppressed by the partial sodium-dependent neutral amino acid transporter (SNAT) antagonist MeAIB and augmented by the Cl- current blocker DCPIB. Electrophysiological recordings showed that addition of glycine activates a Cl- current under isotonic conditions resembling features of the swelling-activated Cl- current (IClswell). The cell membrane potential (Vmem) displayed a distinctive time course after glycine application; initially, glycine evoked a rapid depolarization mediated by Na+-coupled glycine uptake via SNAT, followed by a further gradual depolarization, which was fully suppressed by DCPIB. Interestingly, glycine significantly increased migration of BV-2 cells, which was suppressed by MeAIB, suggesting that SNAT is involved in the migration process of microglial cells. Conclusion: We conclude that glycine acts as a chemoattractant for microglial cells presumably by a cell volume-dependent mechanism involving SNAT-mediated cell swelling.https://www.karger.com/Article/FullText/494646Swelling-dependent chloride currentCl-IClswellVRACVSORCell volume regulationGlycineNeutral amino acid transporterMigrationMicroglia |
| spellingShingle | Michael Kittl Heidemarie Dobias Marlena Beyreis Tobias Kiesslich Christian Mayr Martin Gaisberger Markus Ritter Hubert H. Kerschbaum Martin Jakab Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling Cellular Physiology and Biochemistry Swelling-dependent chloride current Cl- IClswell VRAC VSOR Cell volume regulation Glycine Neutral amino acid transporter Migration Microglia |
| title | Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling |
| title_full | Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling |
| title_fullStr | Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling |
| title_full_unstemmed | Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling |
| title_short | Glycine Induces Migration of Microglial BV-2 Cells via SNAT-Mediated Cell Swelling |
| title_sort | glycine induces migration of microglial bv 2 cells via snat mediated cell swelling |
| topic | Swelling-dependent chloride current Cl- IClswell VRAC VSOR Cell volume regulation Glycine Neutral amino acid transporter Migration Microglia |
| url | https://www.karger.com/Article/FullText/494646 |
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