Glycine modulates membrane potential, cell volume, and phagocytosis in murine microglia

Amino Acids. 2014 Aug;46(8):1907-17. doi: 10.1007/s00726-014-1745-8. Epub 2014 Apr 24.

Abstract

Phagocytes form engulfment pseudopodia at the contact area with their target particle by a process resembling cell volume (CV) regulatory mechanisms. We evaluated whether the osmoregulatory active neutral amino acid glycine, which contributes to CV regulation via activation of sodium-dependent neutral amino acid transporters (SNATs) improves phagocytosis in isotonic and hypertonic conditions in the murine microglial cell line BV-2 and primary microglial cells (pMG). In BV-2 cells and pMG, RT-PCR analysis revealed expression of SNATs (Slc38a1, Slc38a2), but not of GlyRs (Glra1-4). In BV-2 cells, glycine (5 mM) led to a rapid Na(+)-dependent depolarization of membrane potential (V mem). Furthermore, glycine increased CV by about 9%. Visualizing of phagocytosis of polystyrene microspheres by scanning electron microscopy revealed that glycine (1 mM) increased the number of BV-2 cells containing at least one microsphere by about 13%. Glycine-dependent increase in phagocytosis was suppressed by the SNAT inhibitor α-(methylamino)isobutyric acid (MeAIB), by replacing extracellular Na(+) with choline, and under hypertonic conditions, but not by the GlyR antagonist strychnine or the GlyR agonist taurine. Interestingly, hypertonicity-induced suppression of phagocytosis was rescued by glycine. These findings demonstrate that glycine increases phagocytosis in iso- and hypertonic conditions by activation of SNATs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Transport System A / antagonists & inhibitors
  • Amino Acid Transport System A / biosynthesis
  • Amino Acid Transport System A / genetics*
  • Animals
  • Cell Size / drug effects
  • Cells, Cultured
  • Choline / pharmacology
  • Glycine / pharmacology*
  • Glycine Agents / pharmacology
  • Hypertonic Solutions
  • Membrane Potentials / drug effects*
  • Mice
  • Mice, Inbred C57BL
  • Microglia / cytology
  • Microspheres
  • Phagocytosis / drug effects*
  • Polystyrenes
  • Primary Cell Culture
  • RNA, Messenger / biosynthesis
  • Receptors, Glycine / agonists
  • Receptors, Glycine / antagonists & inhibitors
  • Receptors, Glycine / biosynthesis
  • Strychnine / pharmacology
  • Taurine / pharmacology
  • beta-Alanine / analogs & derivatives
  • beta-Alanine / pharmacology

Substances

  • Amino Acid Transport System A
  • Glra1 protein, mouse
  • Glycine Agents
  • Hypertonic Solutions
  • Polystyrenes
  • RNA, Messenger
  • Receptors, Glycine
  • Slc38a1 protein, mouse
  • Slc38a2 protein, mouse
  • beta-Alanine
  • 2,2-dimethyl-beta-alanine
  • Taurine
  • Strychnine
  • Choline
  • Glycine