Multiplexed cytokine protein expression profiles from spreading depression in hippocampal organotypic cultures

Journal of Cerebral Blood Flow and Metabolism : Official Journal of the International Society of Cerebral Blood Flow and Metabolism
Phillip E KunklerRichard P Kraig

Abstract

Cytokines are involved in ischemic tolerance, including that triggered by spreading depression (SD), yet their roles in neuroprotection remain incompletely defined. The latter may stem from the pleiotropic nature of these signaling molecules whose complexities for interaction might be better deciphered through simultaneous measurement of multiple targeted proteins. Accordingly, the authors used microsphere-based flow cytometric immunoassays and hippocampal organotypic cultures (HOTCs) to characterize the magnitude, time course, and diversity of cytokine (interleukin [IL] 1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-10, granulocyte-macrophage colony-stimulating factor [GM-CSF], interferon-gamma [IFN-gamma], and tumor necrosis factor-alpha [TNF-alpha]) response to SD. GM-CSF was not detected in HOTCs or media. However, SD triggered a significant, generalized increase in seven cytokines evident in HOTCs 6 hours later, with the remaining cytokine, IL-1beta, becoming significantly different at 1 and 3 days. Additionally, these changes extended to include surrounding media for IL-6 and TNF-alpha by 1 and 3 days. This increase was localized to microglia via immunostaining for IL-1alpha, IL-1beta, and interferon-y. IL-10, although significan...Continue Reading

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Citations

Feb 6, 2013·BMC Medicine·Else EisingMichel D Ferrari
May 5, 2011·PloS One·Yelena Y GrinbergRichard P Kraig
Apr 15, 2005·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Phillip E KunklerRichard P Kraig
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Aug 28, 2021·International Journal of Molecular Sciences·Gaku YamanakaHisashi Kawashima

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Methods Mentioned

BETA
Assay
dot blots
Protein Array
light microscopy
enzyme-linked immunosorbent assay
enzyme-linked immunosorbent assays

Software Mentioned

SPSS
CorelDraw
Axoscope
SigmaPlot
Plex Manager
Photoshop
Image ProPlus
SigmaStat
Bio
Excel

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