Abstract
Experiments in rodents revealed neuropeptide S (NPS) to constitute a potential novel treatment option for anxiety diseases such as panic and post-traumatic stress disorder. However, both its cerebral target sites and the molecular underpinnings of NPS-mediated effects still remain elusive. By administration of fluorophore-conjugated NPS, we pinpointed NPS target neurons in distinct regions throughout the entire brain. We demonstrated their functional relevance in the hippocampus. In the CA1 region, NPS modulates synaptic transmission and plasticity. NPS is taken up into NPS receptor-expressing neurons by internalization of the receptor–ligand complex as we confirmed by subsequent cell culture studies. Furthermore, we tracked internalization of intranasally applied NPS at the single-neuron level and additionally demonstrate that it is delivered into the mouse brain without losing its anxiolytic properties. Finally, we show that NPS differentially modulates the expression of proteins of the glutamatergic system involved inter alia in synaptic plasticity. These results not only enlighten the path of NPS in the brain, but also establish a non-invasive method for NPS administration in mice, thus strongly encouraging translation into a novel therapeutic approach for pathological anxiety in humans.
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Acknowledgements
We thank Carsten T Wotjak for important feedback on the microscopy data and for crucial help with the statistical analysis. We also thank Christoph Thoeringer and Theo Rein for fruitful discussion on the confocal microscopy images; Markus Nussbaumer for technical assistance in performing the behavioral tests; Bozidar Novak for technical assistance in performing the immunoblots, for cloning EGFP-NPSR, and for performing real-time PCRs; Christine Huber for technical assistance in performing the immunoblots; and Nancy Xin Ru Wang for performing the RNA extraction and real-time PCRs. We thank Andreas Sailer from Novartis for providing us with (R)-SHA 68, and Benno Pütz and Tonia Ludwig for most important advice on the statistical analysis. The research was supported in part by the Horst Kübler-Foundation, Bad Ragaz. The Horst Kübler-Foundation had no further role in study design, in the collection, analysis, and interpretation of the data; in the writing of the report; and in the decision to submit the paper for publication.
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IA Ionescu, Y-C Yen, F Holsboer, R Landgraf, and U Schmidt declare a conflict of interest due to a patent application on intranasal NPS pending since April 2011. J Dine, DR Buell, L Herrmann, and M Eder declare no conflict of interest.
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Ionescu, I., Dine, J., Yen, YC. et al. Intranasally Administered Neuropeptide S (NPS) Exerts Anxiolytic Effects Following Internalization Into NPS Receptor-Expressing Neurons. Neuropsychopharmacol 37, 1323–1337 (2012). https://doi.org/10.1038/npp.2011.317
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DOI: https://doi.org/10.1038/npp.2011.317
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