There are multiple ways to assess neuroinflammation. You can perform immunohistochemistry by using specific markers for microglial activation (Iba1) , astrocytic response (GFAP), and neuronal loss (NeuN or Fluorojade for dying neurons). You can also look at the levels of various cytokines that are upregulated during inflammation (TNF-alpha, Interleukins, TGF-beta, and so on).
What do you want to see exactly? Glial activation (astrocytes and/or microglial cells), Pro-inflammatory molecules… Furthermore, are your studies in vivo? Let me know!
I think you can use immunohistological section of brain to see macrophage distribution, or a cresyl- violet coloration to visualize the increased cell loss. I recommend you to read this artcle for more explanations: "ANDROGENS MODULATE NEURONAL VULNERABILITY TO KAINATE
LESION" from M. RAMSDEN,1 T. M. SHIN,1 AND C. J. PIKE*
Andrus Gerontology Center, University of Southern California, 3715
McClintock Avenue, Los Angeles, CA 90089-0191, USA
There are multiple ways to assess neuroinflammation. You can perform immunohistochemistry by using specific markers for microglial activation (Iba1) , astrocytic response (GFAP), and neuronal loss (NeuN or Fluorojade for dying neurons). You can also look at the levels of various cytokines that are upregulated during inflammation (TNF-alpha, Interleukins, TGF-beta, and so on).
For neurodegeneration try Flurojade staining and any cell death marker with Neuronal marker Neu N (IHC)...another option is cresyl violet and H& E staining for histological assessment of cell death. To assess the neuroinflammation, first check the stature of pro- inflammatory molecules like TNF-alpha, IL-6 and IL-1 beta. Elevated expression of these inflammatory molecules advocates the state of neuroinflammation. Glial activation is also an important sign of neuroinflammation.
Maria Teresa Viscomi, thank you very much for your interest. My idea is to measure the neuroinflammation and neurodegeneration in vivo (mice, induced by continuous infusion of ICV LPS) with and without the use of a drug with (possible) potential anti-inflammatory. I have a particular interest for the hypothalamus.
Anushruti Ashok, I already read about these tests you have mentioned, but I believe they are most often used to research involving the hippocampus and related areas, but I want to to analyze the hypothalamus, these tests may not serve me (am I wrong?)
Quantify the presence of inflammatory mediators that you mentioned is a great idea! Many thanks for the reply!
Mireille Kameni,
Many thanks for the answer! I also think it's important I evaluate cell loss, as well as proteins related to apoptosis in brain slides.
Thank you! (I'll read the article now!)
Janak Gaire and Aarti Nagayach,
Thanks for the reply.
I had never read about Fluoro-Jade, you've already used? Have any manufacturer preference? There is some difficulty, certain detail or limitation for this method?
Thank you very much!
You could also use micro-PET-scanning to measure micro-glial activation. See this article:
Dobos, N. et al (2012) The role of indoleamine 2,3-dioxygenase in a mouse model of neuroinflammation-induced depression. Journal of Alzheimer’s disease : JAD, 28, 905–15.
Try MRI (magnetic resonance imagine) but you will need someone with experience to analyze the imagines...
The level of tyrosine hydroxylase and I think proinflammatory cytokines.
Immunohistochenistry, ELISA, Western Blotting, RT-PCR can be used for estimation of Neuroinflammatory marker in rat brain. TNF-alpha, IL-6 and IL-1 beta proinflammatory markers while IL-10, IL-4 denotes anti inflammatory markers. Other than this one you can also check GFAP, CD11B,Iba-1 expression to check microglial activation.
Using ELISA, W Blot or IHC, you can assess neuroinflammation by using anti-GFAP (for gliosis) and microglial markers. For better characterization, use anti-cytokines too.
Most of these are for post-mortem measurement. Are there ways (aside from the specialized MRI machines being used for some grants right now) for living HUMANS to have neuroinflammation measured?
André Barretto Vianna, in which context are the inflammatory CNS reactions to be quantified? Along arterial, micro- or venous vessels? in the which tissues? after which kind of "inflammation-provoking" maneuver? in relation to a particular quantifiable insult or kind of tissue-alteration?
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