STIMs and Orai1 regulate cytokine production in spinal astrocytes

Our previous study demonstrated that a store-operated calcium channel (SOCC) inhibitor (YM-58483) has central analgesic effects. However, the cellular and molecular mechanisms of such effects remain to be determined. It is well-known that glial cells play important roles in central sensitization. SO...

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Bibliographic Details
Published in:Journal of neuroinflammation 2016-05, Vol.13 (1), p.126-126, Article 126
Main Authors: Gao, Xinghua, Xia, Jingsheng, Munoz, Frances M, Manners, Melissa T, Pan, Rong, Meucci, Olimpia, Dai, Yue, Hu, Huijuan
Format: Article
Language:English
Subjects:
Anilides - pharmacology
Animals
Animals, Newborn
Astrocytes
Astrocytes - drug effects
Astrocytes - metabolism
Calcium channels
Cell Survival - drug effects
Cell Survival - physiology
Cells, Cultured
Cytokines
Cytokines - biosynthesis
Female
Mice
ORAI1 Protein - antagonists & inhibitors
ORAI1 Protein - physiology
Physiological aspects
Pregnancy
Spinal Cord - drug effects
Spinal Cord - metabolism
Stromal Interaction Molecule 1 - antagonists & inhibitors
Stromal Interaction Molecule 1 - physiology
Stromal Interaction Molecule 2 - antagonists & inhibitors
Stromal Interaction Molecule 2 - physiology
Thiadiazoles - pharmacology
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Summary:Our previous study demonstrated that a store-operated calcium channel (SOCC) inhibitor (YM-58483) has central analgesic effects. However, the cellular and molecular mechanisms of such effects remain to be determined. It is well-known that glial cells play important roles in central sensitization. SOC entry (SOCE) has been implicated in many cell types including cortical astrocytes. However, the role of the SOCC family in the function of astrocytes has not been determined. Here, we thoroughly investigated the expression and the functional significance of SOCCs in spinal astrocytes. Primary cultured astrocytes were prepared from neonatal (P2-P3) CD1 mice. Expressions of mRNAs and proteins were respectively assessed by real-time PCR and Western blot analysis. SOCE was measured using a calcium imaging system. Live-cell STIM1 translocation was detected using a confocal microscope. Cytokine levels were measured by the enzyme-linked immunosorbent assay. We found that the SOCC family is expressed in spinal astrocytes and that depletion of calcium stores from the endoplasmic reticulum by cyclopiazonic acid (CPA) resulted in a large sustained calcium entry, which was blocked by SOCC inhibitors. Using the siRNA knockdown approach, we identified STIM1 and Orai1 as primary components of SOCCs in spinal astrocytes. We also observed thapsigargin (TG)- or CPA-induced puncta formation of STIM1 and Orai1. In addition, activation of SOCCs remarkably promoted TNF-α and IL-6 production in spinal astrocytes, which were greatly attenuated by knockdown of STIM1 or Orai1. Importantly, knockdown of STIM2 and Orai1 dramatically decreased lipopolysaccharide-induced TNF-α and IL-6 production without changing cell viability. This study presents the first evidence that STIM1, STIM2, and Orai1 mediate SOCE and are involved in cytokine production in spinal astrocytes. Our findings provide the basis for future assessment of SOCCs in pain and other central nervous system disorders associated with abnormal astrocyte activities.
ISSN:1742-2094
1742-2094
DOI:10.1186/s12974-016-0594-7