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The ascending median raphe projections are mainly glutamatergic in the mouse forebrain

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Abstract

The median raphe region (MRR) is thought to be serotonergic and plays an important role in the regulation of many cognitive functions. In the hippocampus (HIPP), the MRR exerts a fast excitatory control, partially through glutamatergic transmission, on a subpopulation of GABAergic interneurons that are key regulators of local network activity. However, not all receptors of this connection in the HIPP and in synapses established by MRR in other brain areas are known. Using combined anterograde tracing and immunogold methods, we show that the GluN2A subunit of the NMDA receptor is present in the synapses established by MRR not only in the HIPP, but also in the medial septum (MS) and in the medial prefrontal cortex (mPFC) of the mouse. We estimated similar amounts of NMDA receptors in these synapses established by the MRR and in local adjacent excitatory synapses. Using retrograde tracing and confocal laser scanning microscopy, we found that the majority of the projecting cells of the mouse MRR contain the vesicular glutamate transporter type 3 (vGluT3). Furthermore, using double retrograde tracing, we found that single cells of the MRR can innervate the HIPP and mPFC or the MS and mPFC simultaneously, and these double-projecting cells are also predominantly vGluT3-positive. Our results indicate that the majority of the output of the MRR is glutamatergic and acts through NMDA receptor-containing synapses. This suggests that key forebrain areas receive precisely targeted excitatory input from the MRR, which is able to synchronously modify activity in those regions via individual MRR cells with dual projections.

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Acknowledgments

We thank Emőke Szépné Simon, Katalin Lengyel, Katalin Iványi and Győző Goda for the excellent technical assistance and Dr. Ferenc Mátyás for technical discussions. We thank Dr. Viktor Varga for his comments on the previous version of the manuscript. The authors wish to thank László Barna, the Nikon Microscopy Center at IEM, Nikon Austria GmbH and Auro-Science Consulting Ltd for kindly providing technical support for fluorescent microscopy. This work was supported by the National Institutes of Health (grant number NS030549), National Office for Research and Technology–Hungarian Scientific Research Fund (NKTH-OTKA, grant number CNK77793, K83251) and European Research Council (grant number ERC-2011-ADG-294313, SERRACO). András Szőnyi was supported by the European Union and the State of Hungary, co-financed by the European Social Fund in the framework of TÁMOP 4.2.4. A/1-11-1-2012-0001 “National Excellence Program”. Gabor Nyiri was supported by a János Bolyai Research Scholarship.

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  1. Laboratory of Cerebral Cortex Research, Institute of Experimental Medicine Hungarian Academy of Sciences, Budapest, 1083, Hungary

    András Szőnyi, Márton I. Mayer, Csaba Cserép, Virág T. Takács, Tamás F. Freund & Gábor Nyiri

  2. Department of Anatomy, Hokkaido University School of Medicine, Sapporo, 060-8638, Japan

    Masahiko Watanabe

  3. János Szentágothai Doctoral School of Neurosciences, Semmelweis University, Budapest, 1085, Hungary

    András Szőnyi

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  1. András Szőnyi
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  2. Márton I. Mayer
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Szőnyi, A., Mayer, M.I., Cserép, C. et al. The ascending median raphe projections are mainly glutamatergic in the mouse forebrain. Brain Struct Funct 221, 735–751 (2016). https://doi.org/10.1007/s00429-014-0935-1

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