TNFα levels and macrophages expression reflect an i... [PLoS One. 2013] - PubMed - NCBI

TNFα levels and macrophages expression reflect an i... [PLoS One. 2013] - PubMed - NCBI

PLoS One. 2013;8(1):e52394. doi: 10.1371/journal.pone.0052394. Epub 2013 Jan 11.

TNFα levels and macrophages expression reflect an inflammatory potential of trigeminal ganglia in a mouse model of familial hemiplegic migraine.

Franceschini A, Vilotti S, Ferrari MD, van den Maagdenberg AM, Nistri A, Fabbretti E.

Source

Neuroscience Department, International School for Advanced Studies, Trieste, Italy.

Abstract

Latent changes in trigeminal ganglion structure and function resembling inflammatory conditions may predispose to acute attacks of migraine pain. Here, we investigated whether, in trigeminal sensory ganglia, cytokines such as TNFα might contribute to a local inflammatory phenotype of a transgenic knock-in (KI) mouse model of familial hemiplegic migraine type-1 (FHM-1). To this end, macrophage occurrence and cytokine expression in trigeminal ganglia were compared between wild type (WT) and R192Q mutant Ca(V)2.1 Ca(2+) channel (R192Q KI) mice, a genetic model of FHM-1. Cellular and molecular characterization was performed using a combination of confocal immunohistochemistry and cytokine assays. With respect to WT, R192Q KI trigeminal ganglia were enriched in activated macrophages as suggested by their morphology and immunoreactivity to the markers Iba1, CD11b, and ED1. R192Q KI trigeminal ganglia constitutively expressed higher mRNA levels of IL1β, IL6, IL10 and TNFα cytokines and the MCP-1 chemokine. Consistent with the report that TNFα is a major factor to sensitize trigeminal ganglia, we observed that, following an inflammatory reaction evoked by LPS injection, TNFα expression and macrophage occurrence were significantly higher in R192Q KI ganglia with respect to WT ganglia. Our data suggest that, in KI trigeminal ganglia, the complex cellular and molecular environment could support a new tissue phenotype compatible with a neuroinflammatory profile. We propose that, in FHM patients, this condition might contribute to trigeminal pain pathophysiology through release of soluble mediators, including TNFα, that may modulate the crosstalk between sensory neurons and resident glia, underlying the process of neuronal sensitisation.

PMID:

23326332

[PubMed - indexed for MEDLINE]

PMCID:

PMC3543418

Free PMC Article

Images from this publication.See all images (6)Free text

Figure 5TNFα expression in WT and KI ganglia.

A, Representative confocal microscopy images of WT (top row) or R192Q KI (bottom row) trigeminal ganglion sections immunostained for Iba1 (red) or TNFα(green) in basal condition. Pseudocolor images showing areas of high (yellow) and low (blue) Iba1-TNFα expressing cell co-localization. Color scale was also included. Note TNFα immunostaining detected as spots along perimembrane regions. The larger magnification insets show immunostaining of Iba1-TNFα signal (yellow) in KI rather than WT. Scale bar: 30 µm, for large images; Scale bar: 10 µm for larger magnification insets. B, Histograms quantify the percentage of TNFα immunoreactivity over the total of Iba1 expressing cells in different V1, V2 or V3 trigeminal regions (ROI: 370×370 µm). n = 4 WT and 4 R192Q KI mice; * p<0 .05.="" are="" as="" data="" div="" expressed="" mean="" s.d.="">

TNF%3F Levels and Macrophages Expression Reflect an Inflammatory Potential of Trigeminal Ganglia in a Mouse Model of Familial Hemiplegic Migraine

PLoS One. 2013;8(1):e52394.

Figure 6LPS evoked acute TNFα expression in R192Q KI ganglia.

A, Representative confocal microscopy images of WT (top row) or KI (bottom row) trigeminal ganglion sections immunostained for Iba1 (red) or TNFα(green) after saline (left) or LPS injection (i.p., 5 h; right). LPS evokes TNFα expression in WT and KI after injection. Scale bar: 20 µm. B, C, Histograms quantify the occurrence of Iba1 signal (B) and Iba1-TNFα co-localisation (C) in different V1, V2 or V3 trigeminal ganglion regions from WT or KI mice, after saline or LPS injection (i.p., 5 h). ROI: 370×370 µm. n = 3 WT and 3 R192Q KI mice; * p<0 .05="" 1="" 3="" 5="" and="" are="" as="" changes="" content="" cytokine="" d="" data="" div="" e="" expressed="" fold="" following="" from="" ganglia="" h="" histograms="" i.p.="" in="" increase="" ki="" levels="" lps-injected="" lps-injection="" mg="" mice.="" mice="" mrna="" n="3" or="" p="" pg="" protein="" quantify="" r192q="" respect="" saline-injected="" samples="" tnf="" to="" whole="" with="" wt="">

TNF%3F Levels and Macrophages Expression Reflect an Inflammatory Potential of Trigeminal Ganglia in a Mouse Model of Familial Hemiplegic Migraine

PLoS One. 2013;8(1):e52394.

Figure 1Iba1 immunoreactivity in trigeminal ganglia from in WT and R192Q KI mice.

A, Representative confocal microscopy image of a longitudinal section of mouse trigeminal ganglion immunostained for neuronal β-tubulin III (green), and labeled with DAPI (blue). Discrete distribution of neuronal somata in three histological subdivisions of the trigeminal ganglion (i.e. V1, V2 and V3) is indicated by ellipsoids. Scale bar: 300 µm. B, Representative confocal microscopy images of WT (top row) or R192Q KI (bottom row) trigeminal ganglion sections from different V1, V2 and V3 regions of WT and KI ganglia and immunostained with Iba1 (red). Nuclear signal is also shown (blue). Scale bar: 15 µm. C, Histograms quantify Iba1-positive cells in V1, V2 and V3 regions (ROI: 370 µm×370 µm) of WT and KI trigeminal ganglia. Data were collected in parallel from at least 3 WT and 3 KI mice; * p<0 .01.="" 20="" 30="" 640="" an="" and="" anti-glutamine="" anti-iba1="" antibodies="" are="" average="" bar:="" blue="" bottom="" cells.="" cells="" co-localisation="" confocal="" counted="" d="" dapi="" data="" div="" e="" enriched="" f="" fiber-="" found="" from="" ganglia="" ganglion="" glial="" green="" gs-immunolabeled="" gs-positive="" histograms="" iba1-immunoreactivity="" iba1="" iii-positive="" images="" immunoreactivity="" immunostained="" immunostaining="" in="" ki="" labeled="" m.="" macrophages="" mice.="" mice="" n="3" neuronal-="" neurons="" no="" not="" note="" nuclei="" of="" or="" p="" panels="" peculiar="" per="" positive="" quantify="" r192q="" red="" regions="" respect="" respectively="" roi:="" rois="" row="" samples.="" satellite="" scale="" sections="" show="" shown="" slices="" surrounding="" synthetase="" the="" to="" top="" trigeminal="" tubulin="" typically="" was="" were="" whole="" with="" wt="">

TNF%3F Levels and Macrophages Expression Reflect an Inflammatory Potential of Trigeminal Ganglia in a Mouse Model of Familial Hemiplegic Migraine

PLoS One. 2013;8(1):e52394.