PMID- 25451082
OWN - NLM
STAT- MEDLINE
DCOM- 20150810
LR  - 20181202
IS  - 1873-3913 (Electronic)
IS  - 0898-6568 (Linking)
VI  - 27
IP  - 1
DP  - 2015 Jan
TI  - Regulation of phagocytosis and cytokine secretion by store-operated calcium entry 
      in primary isolated murine microglia.
PG  - 177-86
LID - S0898-6568(14)00349-0 [pii]
LID - 10.1016/j.cellsig.2014.11.003 [doi]
AB  - Microglia are immune effector cells in the central nervous system that 
      participate in tissue repair, inflammatory responses, and neuronal degeneration. 
      The most important signaling factor in the differentiation of immune-active cells 
      after stimulation is the sustained high calcium concentration in the cytosol, 
      which is called store-operated calcium entry (SOCE). Recently, the molecular 
      identity of the store-operated channel (SOC) has revealed that Orai1, Orai2, 
      Orai3, Stim1, and Stim2 constitute the most of SOC. In this study, we demonstrate 
      that Orai1- and Stim1-mediated SOC regulated the phagocytic activity and cytokine 
      release of primary isolated murine microglia. RT-PCR analysis revealed that 
      primary cultured microglia from neonatal ICR mouse brains had Orai1, Orai2, 
      Orai3, and Stim1. To elucidate the role of SOCE in the immune functions of 
      microglia, pharmacological inhibitors or knockdown with Orai1 or Stim1 siRNA was 
      applied, and UDP-induced phagocytic activity and LPS-induced cytokine secretion 
      activity were compared. The pharmacological inhibition and siRNA effect was 
      verified by measuring thapsigargin (TG)-, ATP-, or UDP-activated SOCE Ca2+ influx 
      and proper siRNA-mediated knockdown was verified by western blot analysis. 
      UDP-induced phagocytic activity was inhibited by pharmacological inhibitors of 
      SOCE, such as SKF96365 or 2-APB, and knockdown of Orai1 and Stim1. Cytokine 
      secretion of TNF-alpha and IL-6 by LPS treatment was also inhibited by SKF96365 and 
      knockdown of Orai1 and Stim1. Meanwhile, LPS stimulation-induced NF-kappaB activation 
      was not altered, but NFAT1 activity was attenuated with Stim1 knockdown. These 
      results indicate that SOCE, which was composed of Orais and Stim1, regulates 
      UDP-induced phagocytosis and LPS-stimulated cytokine secretion in microglia.
CI  - Copyright (c) 2014 Elsevier Inc. All rights reserved.
FAU - Heo, Dae Keon
AU  - Heo DK
AD  - Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, 
      Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
FAU - Lim, Hye Min
AU  - Lim HM
AD  - Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, 
      Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
FAU - Nam, Joo Hyun
AU  - Nam JH
AD  - Department of Physiology, Dongguk University College of Medicine, Gyeongju 
      780-714, Republic of Korea; Channelopathy Research Center, Dongguk University 
      College of Medicine, Goyang 410-773, Republic of Korea.
FAU - Lee, Min Goo
AU  - Lee MG
AD  - Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, 
      Yonsei University College of Medicine, Seoul 120-752, Republic of Korea.
FAU - Kim, Joo Young
AU  - Kim JY
AD  - Department of Pharmacology and Brain Korea 21 PLUS Project for Medical Science, 
      Yonsei University College of Medicine, Seoul 120-752, Republic of Korea. 
      Electronic address: jooyoungkim@yuhs.ac.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20141107
PL  - England
TA  - Cell Signal
JT  - Cellular signalling
JID - 8904683
RN  - 0 (Calcium Channels)
RN  - 0 (Cytokines)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
RN  - 0 (NFATC Transcription Factors)
RN  - 0 (ORAI1 Protein)
RN  - 0 (Orai1 protein, mouse)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Stim1 protein, mouse)
RN  - 0 (Stromal Interaction Molecule 1)
RN  - 58-98-0 (Uridine Diphosphate)
RN  - 67526-95-8 (Thapsigargin)
RN  - EC 3.4.21.4 (Trypsin)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Calcium/*metabolism
MH  - Calcium Channels/metabolism
MH  - Cell Separation
MH  - Cells, Cultured
MH  - Cytokines/*metabolism
MH  - Gene Knockdown Techniques
MH  - Lipopolysaccharides/pharmacology
MH  - Mice
MH  - Microglia/drug effects/*metabolism
MH  - NF-kappa B/metabolism
MH  - NFATC Transcription Factors/metabolism
MH  - ORAI1 Protein
MH  - *Phagocytosis/drug effects
MH  - RNA, Small Interfering/metabolism
MH  - Signal Transduction/drug effects
MH  - Stromal Interaction Molecule 1
MH  - Thapsigargin/pharmacology
MH  - Transfection
MH  - Trypsin/metabolism
MH  - Uridine Diphosphate/pharmacology
OTO - NOTNLM
OT  - Cytokine secretion
OT  - Microglia
OT  - Orai1 (CRACM1 or TMEM142A)
OT  - Phagocytosis
OT  - Stim1
OT  - Store-operated calcium entry (SOCE)
EDAT- 2014/12/03 06:00
MHDA- 2015/08/11 06:00
CRDT- 2014/12/03 06:00
PHST- 2014/09/19 00:00 [received]
PHST- 2014/10/17 00:00 [revised]
PHST- 2014/11/03 00:00 [accepted]
PHST- 2014/12/03 06:00 [entrez]
PHST- 2014/12/03 06:00 [pubmed]
PHST- 2015/08/11 06:00 [medline]
AID - S0898-6568(14)00349-0 [pii]
AID - 10.1016/j.cellsig.2014.11.003 [doi]
PST - ppublish
SO  - Cell Signal. 2015 Jan;27(1):177-86. doi: 10.1016/j.cellsig.2014.11.003. Epub 2014 
      Nov 7.