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Effects of LPS on P2X3 receptors of trigeminal sensory neurons and macrophages from mice expressing the R192Q Cacna1a gene mutation of familial hemiplegic migraine-1

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Abstract

A knockin (KI) mouse model with the R192Q missense mutation in the Cacna1a gene commonly detected in familial hemiplegic migraine was used to study whether trigeminal ganglia showed a basal inflammatory profile that could be further enhanced by the lipopolysaccharide (LPS) toxin. Adenosine-5′-triphosphate (ATP)-gated purinergic ionotropic receptor 3 (P2X3) currents expressed by the large majority of trigeminal sensory neurons were taken as functional readout. Cultured R192Q KI trigeminal ganglia showed higher number of active macrophages, basal release of tumor necrosis factor alpha (TNFα), and larger P2X3 receptor currents with respect to wild type (WT) cells. After 5 h application of LPS in vitro, both WT and R192Q KI cultures demonstrated significant increase in macrophage activation, very large rise in TNFα mRNA content, and ambient protein levels together with fall in TNFα precursor, suggesting potent release of this inflammatory mediator. Notwithstanding the unchanged expression of P2X3 receptor protein in WT or R192Q KI cultures, LPS evoked a large rise in WT neuronal currents that recovered faster from desensitization. Basal R192Q KI currents were larger than WT ones and could not be further augmented by LPS. These data suggest that KI cultures had a basal neuroinflammatory profile that might facilitate the release of endogenous mediators (including ATP) to activate constitutively hyperfunctional P2X3 receptors and amplify nociceptive signaling by trigeminal sensory neurons.

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Fig. 1
Fig. 2

Abbreviations

α,β-meATP:

α,β-methyleneadenosine 5′-triphosphate

ATP:

Adenosine-5′-triphosphate

AU:

Arbitrary unit

BCA:

Bicinchoninic acid

CaV2.1:

Voltage-activated calcium channel 2.1

DRG:

Dorsal root ganglion

FHM-1:

Familial hemiplegic migraine type 1

Iba1:

Ionized calcium binding adaptor molecule 1

KI:

Knockin

LPS:

Lipopolysaccharide

P2X3:

Purinergic ionotropic receptor 3

PCR:

Polymerase chain reaction

SEM:

Standard error of the mean

TNFα:

Tumor necrosis factor alpha

WT:

Wild type

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Acknowledgments

This work was supported by the Telethon Foundation (grant GGP10082 to AN), the Cariplo Foundation (2011-0505 to AN), and by ARRS grant J3-2376-1540 (to EF) and a grant from the Centre for Medical Systems Biology within the framework of the Netherlands Genomics Initiative/Netherlands Organization for Scientific Research (NWO).

Conflict of interests

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Neuroscience and Italian Institute of Technology Unit, International School for Advanced Studies (SISSA), Via Bonomea 265, 34136, Trieste, Italy

    Alessia Franceschini, Swathi K. Hullugundi, Andrea Nistri & Elsa Fabbretti

  2. Departments of Human Genetics & Neurology, Leiden University Medical Centre, 2300 RC, Leiden, The Netherlands

    Arn M. J. M. van den Maagdenberg

  3. University of Nova Gorica, Nova Gorica, SI-5000, Slovenia

    Elsa Fabbretti

Authors
  1. Alessia Franceschini
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  2. Swathi K. Hullugundi
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  3. Arn M. J. M. van den Maagdenberg
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  4. Andrea Nistri
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  5. Elsa Fabbretti
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Corresponding author

Correspondence to Andrea Nistri.

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Franceschini, A., Hullugundi, S.K., van den Maagdenberg, A.M.J.M. et al. Effects of LPS on P2X3 receptors of trigeminal sensory neurons and macrophages from mice expressing the R192Q Cacna1a gene mutation of familial hemiplegic migraine-1. Purinergic Signalling 9, 7–13 (2013). https://doi.org/10.1007/s11302-012-9328-1

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  • DOI: https://doi.org/10.1007/s11302-012-9328-1

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