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Verification of γ-Amino-Butyric Acid (GABA) Signaling System Components in Periodontal Ligament Cells In Vivo and In Vitro

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Abstract

CNS key neurotransmitter γ-amino-butyric acid (GABA) and its signaling components are likewise detectable in non-neuronal tissues displaying inter alia immunomodulatory functions. This study aimed at identifying potential glutamate decarboxylase (GAD)65 and GABA receptor expression in periodontal ligament (PDL) cells in vivo and in vitro, with particular regard to inflammation and mechanical loading. Gene expression was analyzed in human PDL cells at rest or in response to IL-1ß (5 ng/ml) or TNFα (5 ng/ml) challenge via qRT-PCR. Western blot determined constitutive receptor expression, and confocal laser scanning fluorescence microscopy visualized expression changes induced by inflammation. ELISA quantified GAD65 release. Immunocytochemistry was performed for GABA component detection in vitro on mechanically loaded PDL cells, and in vivo on rat upper jaw biopsies with mechanically induced root resorptions. Statistical significance was set at p < 0.05. GABAB1, GABAB2, GABAA1, and GABAA3 were ubiquitously expressed both on gene and protein level. GABAA2 and GAD65 were undetectable in resting cells, but induced by inflammation. GABAB1 exhibited the highest basal gene expression (6.97 % ± 0.16). IL-1ß markedly increased GABAB2 on a transcriptional (57.28-fold ± 12.40) and protein level seen via fluorescence microscopy. TNFα-stimulated PDL cells released GAD65 (3.68 pg/ml ± 0.17 after 24 h, 5.77 pg/ml ± 0.65 after 48 h). Immunocytochemistry revealed GAD65 expression in mechanically loaded PDL cells. In vivo, GABA components were varyingly expressed in an inflammatory periodontal environment. PDL cells differentially express GABA signaling components and secrete GAD65. Inflammation and mechanical loading regulate these neurotransmitter molecules, which are also detectable in vivo and are potentially involved in periodontal pathophysiology.

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Acknowledgments

We thank D. Nguyen and D. Stephens from the Forsyth Institute as much as I. Bay-Müler K. Reifenrath and S. van Dyk from the Department of Orthodontics, University of Bonn for technical support. This research received Grant Support by the Deutsche Gesellschaft für Kieferorthopädie (DGKFO) and NIH/NIDCR Grant DE020906.

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Authors and Affiliations

  1. Department of Orthodontics, Medical Faculty, University of Bonn, Welschnonnenstr. 17, 53111, Bonn, Germany

    Anna Konermann & Andreas Jäger

  2. Forsyth Institute, 245 First St, Cambridge, MA, 02142, USA

    Alpdogan Kantarci, Steven Wilbert & Thomas Van Dyke

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  1. Anna Konermann
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  2. Alpdogan Kantarci
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  3. Steven Wilbert
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  5. Andreas Jäger
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Correspondence to Anna Konermann.

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Konermann, A., Kantarci, A., Wilbert, S. et al. Verification of γ-Amino-Butyric Acid (GABA) Signaling System Components in Periodontal Ligament Cells In Vivo and In Vitro. Cell Mol Neurobiol 36, 1353–1363 (2016). https://doi.org/10.1007/s10571-016-0335-6

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  • DOI: https://doi.org/10.1007/s10571-016-0335-6

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