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  • Phospholipid localization implies microglial morphology and function via Cdc42 in vitro.

Phospholipid localization implies microglial morphology and function via Cdc42 in vitro.

Glia (2017-02-10)
Kyohei Tokizane, Hiroyuki Konishi, Kumiko Makide, Hiroki Kawana, Shinichi Nakamuta, Kozo Kaibuchi, Tomohiko Ohwada, Junken Aoki, Hiroshi Kiyama
ABSTRACT

Under a quiescent state, microglia exhibit a ramified shape, rather than the amoeboid-like morphology following injury or inflammation. The manipulation of microglial morphology in vitro has not been very successful, which has impeded the progress of microglial studies. We demonstrate that lysophosphatidylserine (LysoPS), a kind of lysophospholipids, rapidly and substantially alters the morphology of primary cultured microglia to an in vivo-like ramified shape in a receptor independent manner. This mechanism is mediated by Cdc42 activity. LysoPS is incorporated into the plasma membrane and converted to phosphatidylserine (PS) via the Lands' cycle. The accumulated PS on the membrane recruits Cdc42. Both Cdc42 and PS colocalize predominantly in primary and secondary processes, but not in peripheral branches or tips of microglia. Along with the morphological changes LysoPS suppresses inflammatory cytokine production and NF-kB activity. The present study provides a tool to manipulate a microglial phenotype from an amoeboid to a fully ramified in vitro, which certainly contributes to studies exploring microglial physiology and pathology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
ML 141, ≥98% (HPLC)
Sigma-Aldrich
U0126, U0126, CAS 109511-58-2, is a potent and specific inhibitor of MEK1 (IC₅₀ = 72 nM) and MEK2 (IC₅₀ = 58 nM). The inhibition is noncompetitive with respect to both ATP and ERK.
Sigma-Aldrich
LY 294002, LY294002, CAS 154447-36-6, is a cell-permeable, potent, reversible, and specific inhibitor of PI 3-kinase ((IC₅₀ = 1.4 µM). Acts on the ATP-binding site.
Sigma-Aldrich
KT5720, A potent, specific, cell-permeable, reversible and ATP-competitive inhibitor of protein kinase A (Ki = 56 nM) that is prepared by a chemical modification of K-252a.