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SNAP-25 is abundantly expressed in enteric neuronal networks and upregulated by the neurotrophic factor GDNF

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Abstract

Control of intestinal motility requires an intact enteric neurotransmission. Synaptosomal-associated protein 25 (SNAP-25) is an essential component of the synaptic vesicle fusion machinery. The aim of the study was to investigate the localization and expression of SNAP-25 in the human intestine and cultured enteric neurons and to assess its regulation by the neurotrophic factor glial cell line-derived neurotrophic factor (GDNF). SNAP-25 expression and distribution were analyzed in GDNF-stimulated enteric nerve cell cultures, and synaptic vesicles were evaluated by scanning and transmission electron microscopy. Human colonic specimens were processed for site-specific SNAP-25 gene expression analysis and SNAP-25 immunohistochemistry including dual-labeling with the pan-neuronal marker PGP 9.5. Additionally, gene expression levels and distributional patterns of SNAP-25 were analyzed in colonic specimens of patients with diverticular disease (DD). GDNF-treated enteric nerve cell cultures showed abundant expression of SNAP-25 and exhibited granular staining corresponding to synaptic vesicles. SNAP-25 gene expression was detected in all colonic layers and isolated myenteric ganglia. SNAP-25 co-localized with PGP 9.5 in submucosal and myenteric ganglia and intramuscular nerve fibers. In patients with DD, both SNAP-25 mRNA expression and immunoreactive profiles were decreased compared to controls. GDNF-induced growth and differentiation of cultured enteric neurons is paralleled by increased expression of SNAP-25 and formation of synaptic vesicles reflecting enhanced synaptogenesis. The expression of SNAP-25 within the human enteric nervous system and its downregulation in DD suggest an essential role in enteric neurotransmission and render SNAP-25 as a marker for impaired synaptic plasticity in enteric neuropathies underlying intestinal motility disorders.

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Abbreviations

CNS:

Central nervous system

DD:

Diverticular disease

ENS:

Enteric nervous system

GABA:

γ-Aminobutyric acid

GDNF:

Glial cell line-derived neurotropic factor

GFRα1:

GDNF family receptor alpha 1

GI:

Gastrointestinal

HSCR:

Hirschsprung’s disease

IBS:

Irritable bowel syndrome

LMD:

Laser capture microdissection

qPCR:

Quantitative PCR

RET:

Rearranged during transfection

SNAP-25:

Synaptosome-associated protein 25 kDa

SNARE:

Soluble N-ethylmaleimide-sensitive factor attachment receptor

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Acknowledgments

The authors thank Karin Stengel, Inka Geurink, Miriam Lemmer, Frank Lichte, Bettina Facompré and Clemens Franke (Institute of Anatomy, Christian-Albrechts-University of Kiel) for their excellent technical assistance. This work was supported by research grants from the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG WE 2366/4-3) and the Faculty of Medicine, University of Kiel (F347022). The funding sources have no role in study design, management of data and writing of the paper.

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The authors disclose no competing interests.

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    Authors and Affiliations

    1. Institute of Anatomy, Christian-Albrechts-University of Kiel, Otto-Hahn-Platz 8, 24118, Kiel, Germany

      M. Barrenschee, M. Böttner, J. Harde, C. Lange, F. Cossais & T. Wedel

    2. Department of Pathology, Städtisches Krankenhaus Kiel, Kiel, Germany

      M. Ebsen

    3. Department of Surgery, Städtisches Krankenhaus Kiel, Kiel, Germany

      I. Vogel

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    1. M. Barrenschee
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    Correspondence to M. Barrenschee.

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    M. Barrenschee and M. Böttner have contributed equally to this work.

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    Barrenschee, M., Böttner, M., Harde, J. et al. SNAP-25 is abundantly expressed in enteric neuronal networks and upregulated by the neurotrophic factor GDNF. Histochem Cell Biol 143, 611–623 (2015). https://doi.org/10.1007/s00418-015-1310-x

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