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Co-localization of Gamma-Aminobutyric Acid and Glutamate in Neurons of the Spider Central Nervous System

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Abstract

Spider sensory neurons with cell bodies close to various sensory organs are innervated by putative efferent axons from the central nervous system (CNS). Light and electronmicroscopic imaging of immunolabeled neurons has demonstrated that neurotransmitters present at peripheral synapses include γ-aminobutyric acid (GABA), glutamate and octopamine. Moreover, electrophysiological studies show that these neurotransmitters modulate the sensitivity of peripheral sensory neurons. Here, we undertook immunocytochemical investigations to characterize GABA and glutamate-immunoreactive neurons in three-dimensional reconstructions of the spider CNS. We document that both neurotransmitters are abundant in morphologically distinct neurons throughout the CNS. Labeling for the vesicular transporters, VGAT for GABA and VGLUT for glutamate, showed corresponding patterns, supporting the specificity of antibody binding. Whereas some neurons displayed strong immunolabeling, others were only weakly labeled. Double labeling showed that a subpopulation of weakly labeled neurons present in all ganglia expresses both GABA and glutamate. Double labeled, strongly and weakly labeled GABA and glutamate immunoreactive axons were also observed in the periphery along muscle fibers and peripheral sensory neurons. Electron microscopic investigations showed presynaptic profiles of various diameters with mixed vesicle populations innervating muscle tissue as well as sensory neurons. Our findings provide evidence that: (1) sensory neurons and muscle fibers are innervated by morphologically distinct, centrally located GABA- and glutamate immunoreactive neurons; (2) a subpopulation of these neurons may co-release both neurotransmitters; and (3) sensory neurons and muscles are innervated by all of these neurochemically and morphologically distinct types of neurons. The biochemical diversity of presynaptic innervation may contribute to how spiders filter natural stimuli and coordinate appropriate response patterns.

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Acknowledgments

The authors thank Dr. Roger Croll for the anti-GAT antibody, helpful discussions, suggestions and general support. We are grateful to Dr. Hermann Aberle (University Düsseldorf) for the anti-DVGLUT antibody. We thank Dr. Andrew French for the transcriptome search of Cupiennius VGAT and VGLUT proteins. We also thank Drs. Thomas Trappenberg, Friedrich Barth, Ernst-August Seyfarth and Alan Fine for support and helpful discussions. This study was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC), Discovery grants RGPIN-2014-05565 to P.H.T., and A-0000065 and Accelerator Supplement 429437 (NSERC, Ottawa) to I.A.M.

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  1. Ruth Fabian-Fine

    Present address: Department of Biology, Saint Michael’s College, One Winooski Park, Colchester, VT, 05439, USA

Authors and Affiliations

  1. Department of Psychology and Neuroscience, Life Sciences Centre, Dalhousie University, 1355 Oxford Street, P.O. Box 15000, Halifax, Nova Scotia, Canada, B3H 4R2

    Ruth Fabian-Fine & Ian A. Meinertzhagen

  2. Department of Physiology and Biophysics, Dalhousie University, PO Box 15000, Halifax, Nova Scotia, Canada, B3H 4R2

    Shannon Meisner & Päivi H. Torkkeli

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  1. Ruth Fabian-Fine
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  4. Ian A. Meinertzhagen
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Correspondence to Ruth Fabian-Fine.

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Fabian-Fine, R., Meisner, S., Torkkeli, P.H. et al. Co-localization of Gamma-Aminobutyric Acid and Glutamate in Neurons of the Spider Central Nervous System. Cell Tissue Res 362, 461–479 (2015). https://doi.org/10.1007/s00441-015-2241-5

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