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Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014
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The L-type voltage-gated calcium channel modulates microglial pro-inflammatory activity
J F Espinosa-Parrilla1, M Martínez-Moreno1, X Gasull2
1Unitat de Bioquímica i Biologia Molecular, Facultat de Medicina, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Barcelona, Spain.
Molecular and Cellular Neurosciences
|December 16, 2014
Summary
Activated microglia express L-type voltage-gated calcium channels (VGCCs), influencing inflammatory responses. Blocking these channels reduced cytokine release but not phagocytosis, suggesting a specific role in neuroinflammation.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are key immune cells in the central nervous system (CNS).
- Under pathological conditions, microglia become reactive, releasing pro-inflammatory and neurotoxic factors.
- The role of L-type voltage-gated calcium channels (VGCCs) in microglial activation is not fully understood.
Purpose of the Study:
- To investigate L-type VGCC expression in microglia during neurodegeneration.
- To determine the functional role of L-type VGCCs in microglial cytokine release and phagocytosis.
Main Methods:
- Rat model of N-methyl-D-aspartate-induced hippocampal neurodegeneration.
- Double immunohistochemistry and confocal microscopy to detect L-type VGCC in microglia.
- BV2 microglia cell line used for functional assays.
- Reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunocytochemistry to identify VGCC subunits.
- Calcium imaging and pharmacological manipulation (Bay k8644, nifedipine) to assess channel function and effects on cytokine release and phagocytosis.
Main Results:
- Activated microglia in vivo express L-type VGCC.
- BV2 microglia express CaV1.2 and CaV1.3 subunits of L-type VGCC.
- KCl-induced calcium entry in BV2 cells was modulated by Bay k8644 and nifedipine.
- Nifedipine treatment significantly reduced TNF-α and NO release by 40% but did not affect BV2 cell viability or phagocytic capacity.
Conclusions:
- L-type VGCCs are expressed in activated microglia.
- L-type VGCCs play a specific role in regulating microglial secretory activity, particularly cytokine release.
- Targeting L-type VGCCs may offer a strategy to modulate neuroinflammation without impairing essential microglial functions like phagocytosis.
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