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NCX1 expression and functional activity increase in microglia invading the infarct core.

Francesca Boscia1, Rosaria Gala, Anna Pannaccione

  • 1Division of Pharmacology, Department of Neuroscience, School of Medicine, Federico II, University of Naples, Naples, Italy.

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Sodium-calcium exchanger NCX1 is upregulated in microglia after stroke. This enhanced NCX1 expression and activity in microglial cells plays a key role in the post-ischemic brain.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Ischemic Stroke Research

Background:

  • The sodium-calcium exchanger NCX1 is crucial for maintaining intracellular sodium and calcium levels, especially during anoxic conditions.
  • Limited information exists regarding NCX1 expression and activity in microglial cells following ischemic events.

Purpose of the Study:

  • To investigate the expression and activity of NCX1 protein in microglial cells within the peri-infarct and core regions after permanent middle cerebral artery occlusion (MCAO).
  • To determine the functional role of NCX1 in microglial cells under ischemic and hypoxic conditions.

Main Methods:

  • Western blotting, patch-clamp electrophysiology, single-cell microfluorometry, immunohistochemistry, and confocal microscopy were employed.
  • NCX1 expression and activity were assessed in primary microglia from adult rat brains post-MCAO and in cultured microglia under in vitro hypoxia.
  • NCX1 gene silencing was used to evaluate its specific role in hypoxia-induced calcium changes.

Main Results:

  • NCX1 protein expression increased in microglial cells within the infarct core following MCAO, particularly in cells invading the core region.
  • Primary and BV2 microglial cells from ischemic/hypoxic conditions showed significantly enhanced NCX1 expression and reverse mode activity.
  • NCX1 silencing prevented the hypoxia-induced increase in intracellular calcium in microglial cells, while NCX2 and NCX3 were downregulated.

Conclusions:

  • The observed upregulation of NCX1 expression and activity in microglia after MCAO suggests a significant role for NCX1 in modulating microglial function in the post-ischemic brain.
  • NCX1 is a critical regulator of intracellular calcium homeostasis in microglia during ischemic conditions.