Forschungsgruppenleiter: Rolf Sprengel
Neuropsychiatric disorders are frequently inherited, and in many cases result from spontaneous mutations. Studies with patients have identified many defects in the genes for proteins that play key roles in fast neurotransmission and in brain development, in particular proteins found in excitatory neurons. These are, on the one hand, ion channels activated by the neurotransmitter glutamate and, on the other hand, scaffolding proteins that organize the topographic arrangement and intracellular signaling cascades in response to activation of these ion channels. The latter thus determine the efficiency and the quality of the glutamatergic signal transmission. Our research aims to identify gene markers for diagnostic purposes and to understand the consequences of the defect as a basis for therapeutic approaches. Our focus is on defects associated with schizophrenia, cognition problems and autism. We generate and analyze numerous genetically manipulated mouse models with abnormal cognitive and social behaviors, and try to rescue behavioral impairments by drug treatment. Our genetically or virally manipulated mice have been shown to provide reliable and useful models of schizophrenia and depression. Other mouse lines were more suitable for the analysis of physiological mechanisms underlying short and long-term memory. Over-expression of the structural protein SHANK2 allowed us to show that genetically-related ASD-like behavior in mice can be cured by repairing the gene defect in adult mice.(Barkus, C., et al. Neuropharmacology 62, 1263-1272 (2012); Fitzgerald, P.J., et al. Neurobiol. Dis. 40, 608-621 (2010); Inta, D., et al. Neurosci. Biobehav. Rev. 34, 285-294 (2010); Sanderson, D.J., et al. Prog. Brain Res. 169, 159-178 (2008); Wiedholz, L.M., et al. Mol. Psychiatry 13, 631-640 (2008); Bannerman, D.M., et al. Nat. Neurosci. 15, 1153-1159 (2012); Shimshek, D.R., et al. J. Neuroscience 26, 8428-8440 (2006); Zamanillo, D., et al. Science 284, 1805-1811 (1999)).
The role of the glutamate-gated ion channels and the postsynaptic organizer: SHANK2 in schizophrenia, cognition and autism/attention deficits
Two types of glutamate receptors have been known for many years. The form that is activated by AMPA was recognized as an essential glutamate-gated ion channel involved in synaptic transmission. In 1999 we showed that it is also involved in memory formation. In our recent work we have solidified our 20 year-old hypothesis (Zamanillo, D., et al. Science 284, 1805-1811 (1999); Reisel, D. et al., Nat. Neurosci. 5(9), 868-873 (2002)) that AMPARs with the GluA1 subunit are essential for short-term memory, also called working memory, but are not required for learning complex associations (Bannerman et al., 2018; Shimshek et al., 2017). The loss of GluA1-containing AMPARs in a few brain regions was sufficient to impair working memory, and the genetic repair of GluA1 AMPARs in the hippocampus of GluA1 knock-out animals was sufficient to partially restore short-term memory. From this we could also conclude that several brain regions cooperate in a good short-term memory (Freudenberg et al., 2016). In a very recent study, we were able to show directly that the communication between prefrontal cortex and hippocampus, measured in oscillations, is disturbed in animals with poor short-term memory, and this is precisely when this memory is needed to make a correct decision (Bygrave et al., 2019). This allowed us to establish a direct link between schizophrenia, lack of short-term memory and the risk gene GluA1 for bipolar disorders, as we had postulated in our previous work (Weber et al., 2015; Inta et al., 2014; Vogt et al., 2014; Boerner et al., 2017; Sanderson et al., 2017) and as is supported by the poor short-term memory of schizophrenic patients. More importantly, we could attenuate the hyperactivity of our GluA1 deficient mice by haloperidol and cannabidiol (Boerner et al., 2017, Aitta-Aho et al., 2019), both of which are possible candidates for treatment of schizophrenia. In contrast to GluA1 deficient mice, gene-targeted mice which lack an important AMPAR trafficking protein (FRRS1l) showed severe behavioral and learning impairments similar to the handicaps in a young human patient with FRRS1l deficiency (Schwenk et al. 2019).
The N-methyl-D-aspartate receptors (NMDARs)
By contrast, glutamate receptors which are also activated by NMDA were recognized early on as being involved in controlling the formation and the modulation of neuronal networks: the regulated Ca2+ influx through them triggers cellular mechanisms closely associated with learning and memory. In our recent studies we observed neuropsychiatric defects in mice after cell-type-specific NMDAR knock-out pharmacological NMDAR inhibition (Lang et al., 2018; Bygrave et al., 2016), in line with reports on the behavioral alterations in NMDAR hypomorphic mice and with NMDAR point mutations in patients with variable neurological phenotypes (Mohn et al., Cell 98, 427-436 (1999); Endele, S et al., Nat Genet 42, 1021–1026(2010)). In one patient with cognitive impairments and epileptic seizures, the mutation GluN2A(N615K) inhibited the precise NMDARs-guided Ca2+ influx. The analysis of our mice with the analogous mutation GluN2A(N615S) provided first experimental evidence that muscular hypotonia, audiogenic induced seizures (AGS), pronounced hyperactivity, reduced social interaction and selective inability to learn complex spatial associations are directly linked to the GluN2A(N615S) mutation and its inaccurate Ca2+ signaling (Bertocchi et al., 2019, in preperation). We could rescue hyperactivity by amphetamine and by memantine AGS, indicating the acute involvement of disturbed NMDAR signaling in the neurological phenotype (Figure 1). In summary, the global expression of GluN2A(N615S) unravelled several neuronal networks in the forebrain and the brainstem, which are using precise activity-dependent NMDAR Ca2+ signalling to balance conflicting information between functionally connected neuronal networks. In two reviews we discussed our view of the role of NMDARs in the hippocampus in spatial learning (Bannerman et al., 2014; Taylor et al., 2014).
SH3 and multiple ankyrin repeat domain proteins (SHANKs)
Mutations in the three SHANK genes have frequently been identified in patients with Autism Spectrum Disorders (ASD). The huge spectrum of behavioral abnormalities, ranging from schizophrenia to autism or the Asperger syndrome associated with mutations, is reminiscent of the high diversity of phenotypes of 29 SHANK knock-out mouse lines (Eltokhi et al., 2018). Furthermore, we showed that the sex hormones result in strong gender-dependent differences in the expression of Shank genes (Berkel et al., 2018). Using strong and conditional overexpression of the SHANK2A and the SHANK(R462X) truncation in excitatory forebrain neurons during and after development (Figure 2) we found that the social component of the ASD-like behaviour could be erased when we switched off the SHANK2A overexpression in adult mice, demonstrating that the social component of ASD can be restored by therapeutic treatment but not other components such as hyperactivity, anxiety and cognitive dysfunctions. In both lines the hyperactivity could be attenuated by amphetamine, indicating an increase activity of the glutamatergic system (Eltokhi et al., 2019, in preperation).
Development of virus-mediated gene transfer in the mouse brain
We developed viral vectors for tagging specific neuron and astroglia populations with genetic fluorescence or activity indicators or optical and pharmacological silencers. We provided proof of the long-term expression of virus-delivered secreted peptide hormones in the brain, provided a system for pharmacological long-term silencing of neuronal populations, and evaluated the interaction of different brain regions during the formation and expression of fear (Hasan et al, 2019, Dogbevia et al., 2016; Heinonen et al., 2014; Schwarz et al., 2015; Lissek et al., 2016; Obenhaus et al.,2016; Eliava et al.,2016). We used genetic activity indicators to analyse the kinetics of astrocyte activity in vitro and visualized in vivo the temporal relationship between the cortical spread of glutamate, neuronal and glial activity in an animal model of migraine (Heuser et al., 2018; Enger et al., 2017; Enger et al., 2015; Tang et al., 2015, Enger et al., 2019).
Within the next two years I will complete my research under the collaboration agreement between the Heidelberg University Hospital and the MPImR. Externally funded projects with the Central Institute of Medical Health (CIMH) in Mannheim and the University of Oslo will be continued until June 2021.
Publications (since 2015)
Aitta-aho, T., Maksimovic, M, Dahl, K., Sprengel, R., and Korpi, E. (2019). Attenuation of novelty-induced hyperactivity of Gria1-/- mice by cannabidiol and hippocampal inhibitory chemogenetics. Front. Pharmacol. 10, 309
Bygrave, A. M., Jahans-Price, T., Wolf, A. R., Sprengel, R., Kullman, D. M., Bannerman, D. M., and Kätze, D. (2019). Hippocampal-prefrontal coherence mediates working memory and selective attention at distinct frequency bands and provided a causal link between schizophrenia and its risk gene GRIA1. Transl. Psychiatry 9, 309
Hasan, M. T., Althammer, M., Silva da Gouveia, M., Goyon, S., Eliava, M., Lefevre, A., Kerspern, D., Schimmer, J., Raftogianni, A., Wahis, J., Knobloch-Bollmann, H. S., Tang, Y., Liu, X., Jain, A., Chavant, V., Goumon, Y., Weislogel, J.-M., Hurlemann, R., Herpertz, S. C., Pitzer, C., Darbon, P., Dogbevia, G. K., Bertocchi, I., Larkum, M. E., Sprengel, R., Bading, H., Charlet, A., and Grinevich, V. (2019). Fear memory engram and its plasticity in the hypothalamic oxytocin system. Neuron 103, 133-146
Reinert, J. K., Sonntag, I., Sonntag, H., Sprengel, R., Pelzer, P., Lessle, S., Kaiser, M., and Kuner, T. (2019). RetroLEAP: rAAV-based retrograde trans-synaptic labeling, expression and perturbation. Nat. Methods (under revision)
Schwenk, J., Boudkkazi, S., Kocylowski, M., Brechet, A., Zolles, G., Bus, T., Costa, K., Bildl, W., Sprengel, R., Kulik, A., Roeper, J., Schulte, U. and Fakler, B. (2019). An ER assembly line of AMPA-receptors controls excitatory neurotransmission and its plasticity. Neuron, in press
Ang, G., McKillop, L. E., Purple, R., Blanco-Duque, C., Peirson, S. N., Foster, R. G., Harrison, P. J., Sprengel, R., Davies, K. E., Oliver, P. L., Bannerman, D. M. and Vyazovskiy, V. V. (2018). Absent sleep EEG spindle activity in GluA1 (Gria1) knockout mice: relevance to neuropsychiatric disorders. Transl. Psychiatry 8, 154
Bannerman, D. M., Borchardt, T., Jensen, V., Rozov, A., Haj-Yasein, N. N., Burnashev, N., Zamanillo, D., Bus, T., Grube, I., Adelmann, G., Rawlins, J. N. P. and Sprengel, R. (2018). Somatic accumulation of GluA1-AMPA receptors leads to selective cognitive impairments in mice. Front. Mol. Neurosci. 11, 199
Berkel, S., Eltokhi, A., Frohlich, H., Porras-Gonzalez, D., Rafiullah, R., Sprengel, R. and Rappold, G. A. (2018). Sex hormones regulate SHANK expression. Front. Mol. Neurosci. 11, 337
Eltokhi, A., Rappold, G., and Sprengel, R. (2018). Distinct phenotypes of Shank2 mouse models reflect neuropsychiatric spectrum disorders of human patients wth SHANK2 variants. Front. Mol. Neurosci. 11
Heuser, K., Nome, C. G., Pettersen, K. H., Abjorsbraten, K. S., Jensen, V., Tang, W., Sprengel, R., Tauboll, E, Nagelhus, E. A. and Enger, R. (2018). Ca2+ signals in astrocytes facilitate spread of epileptiform activity. Cereb. Cortex 28, 4036-4048
Lang, E., Mallien, A. S., Vasilescu, A. N., Hefter, D., Luoni, A., Riva, M. A., Borgwardt, S., Sprengel, R., Lang, U. E., Gass, P. and Inta, D. (2018). Molecular and cellular dissection of NMDA receptor subtypes as antidepressant targets. Neurosci. Biobehav. Rev. 84, 352-358
Austen, J. M., Sprengel, R. and Sanderson, D. J. (2017). GluA1 AMPAR subunit deletion reduces the hedonic response to sucrose but leaves satiety and conditioned responses intact. Sci. Rep. 7, 7424
Boerner, T., Bygrave, A. M., Chen, J., Fernando, A., Jackson, S., Barkus, C., Sprengel, R., Seeburg, P. H., Harrison, P. J., Gilmour, G., Bannerman, D. M. and Sanderson, D. J. (2017). The group II metabotropic glutamate receptor agonist LY354740 and the D2 receptor antagonist haloperidol reduce locomotor hyperactivity but fail to rescue spatial working memory in GluA1 knockout mice. Eur. J. Neurosci. 45, 912-921
Broker-Lai, J., Kollewe, A., Schindeldecker, B., Pohle, J., Nguyen Chi, V., Mathar, I., Guzman, R., Schwarz, Y., Lai, A., Weissgerber, P., Schwegler, H., Dietrich, A., Both, M., Sprengel, R., Draguhn, A., Kohr, G., Fakler, B., Flockerzi, V., Bruns, D. and Freichel, M. (2017). Heteromeric channels formed by TRPC1, TRPC4 and TRPC5 define hippocampal synaptic transmission and working memory. EMBO J. 36, 2770-2789
Enger, R., Dukefoss, D. B., Tang, W., Pettersen, K. H., Bjornstad, D. M., Helm, P. J., Jensen V., Sprengel, R., Vervaeke K., Ottersen O. P. and Nagelhus E. A. (2017). Deletion of Aquaporin-4 curtails extracellular glutamate elevation in cortical spreading depression in Awake Mice. Cereb. Cortex 27, 24-33
Gutierrez-Castellanos, N., Da Silva-Matos, C. M., Zhou, K., Canto, C. B., Renner, M. C., Koene, L. M., Ozyildirim, O., Sprengel, R., Kessels, H. W. and De Zeeuw, C. I. (2017). Motor learning requires Purkinje cell synaptic potentiation through activation of AMPA-receptor subunit GluA3. Neuron 93, 409-424
Inta, I., Domonkos, E., Pfeiffer, N., Sprengel, R., Bettendorf, M., Lang, U. E., Inta, D. and Gass, P. (2017). Puberty marks major changes in the hippocampal and cortical c-Fos activation pattern induced by NMDA receptor antagonists. Neuropharmacology 112, 181-187
Kougioumtzidou, E., Shimizu, T., Hamilton, N. B., Tohyama, K., Sprengel, R., Monyer, H., Attwell, D., and Richardson, W. D. (2017). Signalling through AMPA receptors on oligodendrocyte precursors promotes myelination by enhancing oligodendrocyte survival. Elife 6
Lang, E., Mallien, A.S., Vasilescu, A.N., Hefter, D., Luoni, A., Riva, M.A., Borgwardt, S., Sprengel, R., Lang, U.E., Gass, P. and Inta, D. (2017). Molecular and cellular dissection of NMDA receptor subtypes as antidepressant targets. Neurosci. Biobehav. Rev. 84, 352-358
Sanderson, D. J., Lee, A., Sprengel, R., Seeburg, P. H., Harrison, P. J. and Bannerman, D. M. (2017). Altered balance of excitatory and inhibitory learning in a genetically modified mouse model of glutamatergic dysfunction relevant to schizophrenia. Sci. Rep. 7, 1765
Shimshek, D. R., Bus, T., Schupp, B., Jensen, V., Marx, V., Layer, L. E., Kohr, G. and Sprengel, R. (2017). Different forms of AMPA receptor mediated LTP and their correlation to the spatial working memory formation. Front. Mol. Neurosci. 10, 214
Tan, L. L., Pelzer, P., Heinl, C., Tang, W., Gangadharan, V., Flor, H., Sprengel, R., Kuner, T. and Kuner, R. (2017). A pathway from midcingulate cortex to posterior insula gates nociceptive hypersensitivity. Nat. Neurosci. 20, 1591-1601
Bygrave, .A. M., Masiulis, S., Nicholson, E., Berkemann, M., Barkus, C., Sprengel, R., Harrison, P. J., Kullmann, D. M., Bannerman, D. M, and Katzel, D. (2016). Knockout of NMDA-receptors from parvalbumin interneurons sensitizes to schizophrenia-related deficits induced by MK-801. Transl. Psychiatry 6, e778
Dogbevia, G. K., Robetamanith, M., Sprengel, R. and Hasan, M. T. (2016). Flexible, AAV-equipped genetic modules for inducible control of gene expression in mammalian brain. Mol. Ther. Nucleic Acids 5, e309
Eliava, M., Melchior, M., Knobloch-Bollmann, H. S., Wahis, J., da Silva Gouveia, M., Tang, Y., Ciobanu, A. C., Triana Del Rio, R., Roth, L. C., Althammer, F., Chavant, V., Goumon, Y., Gruber, T., Petit-Demouliere, N., Busnelli, M., Chini, B., Tan, L. L., Mitre, M., Froemke, R. C., Chao, M. V., Giese, G., Sprengel, R., Kuner, R., Poisbeau, P., Seeburg, P. H., Stoop, R., Charlet., A. and Grinevich, V. (2016). A new population of parvocellular oxytocin neurons controlling magnocellular neuron activity and inflammatory pain processing. Neuron 89, 1291-1304
Freudenberg, F., Resnik, E., Kolleker, A., Celikel, T., Sprengel, R. and Seeburg, P. H. (2016). Hippocampal GluA1 expression in Gria1-/- mice only partially restores spatial memory performance deficits. Neurobiol Learn. Mem. 135, 83 - 90
Inta, D., Sprengel, R., Borgwardt, S., Lang, U. E. and Gass, P. (2016). The antidepressant effect of ketamine: Mediated by AMPA receptors? Eur. Neuropsychopharmacol. 26, 1692-1693
Lissek, T., Obenhaus, H. A., Ditzel, D. A., Nagai, T., Miyawaki, A., Sprengel, R. and Hasan, M. T. (2016). General anesthetic conditions induce network synchrony and disrupt sensory processing in the cortex. Front. Cell. Neurosci. 10, 64
Obenhaus, H. A., Rozov, A., Bertocchi, I., Tang, W., Kirsch, J., Betz, H. and Sprengel, R. (2016). Causal interrogation of neuronal networks and behavior through virally transduced Ivermectin receptors. Front. Mol. Neurosci. 9, 75
Sprengel, R, and Freudenberg, F (2016). A tribute to Peter H. Seeburg (8.21.1944-8.22.2016) In Memoriam. Neurobiol. Learn. Mem. 136, A1-A2.
Watanabe, Y., Muller, M. K., von Engelhardt, J., Sprengel, R., Seeburg, P. H. and Monyer, H. (2016). Age-dependent degeneration of mature dentate gyrus granule cells following NMDA receptor ablation. Front. Mol. Neurosci. 8, 87
Yang, B., Dormann, C., Vogt, M. A., Sprengel, R., Gass, P. and Inta, D. (2016). Facilitated c-Fos induction in mice deficient for the AMPA receptor-associated protein Ckamp44. Cell. Mol. Neurobiol. 36, 1215 - 1218
Dogbevia, G. K., Marticorena-Alvarez, R., Bausen, M., Sprengel, R. and Hasan, M. T. (2015). Inducible and combinatorial gene manipulation in mouse brain. Front. Cell. Neurosci. 9, 142
Enger, R., Tang, W., Vindedal, G. F., Jensen, V., Johannes Helm, P., Sprengel, R, Looger, L. L. and Nagelhus, E. A. (2015). Dynamics of ionic shifts in cortical spreading depression. Cereb. Cortex 25, 4469-4476
Kiselycznyk, C., Jury, N. J., Halladay, L. R., Nakazawa, K., Mishina, M., Sprengel, R., Grant, S. G., Svenningsson, P. and Holmes, A. (2015). NMDA receptor subunits and associated signaling molecules mediating antidepressant-related effects of NMDA-GluN2B antagonism. Behav. Brain Res. 287, 89-95
Schwarz, M. K., Scherbarth, A., Sprengel, R., Engelhardt, J., Theer, P. and Giese, G. (2015). Fluorescent-protein stabilization and high-resolution imaging of cleared, intact mouse brains. PLoS One 10, e0124650
Steinfeld, R., Herb, J.T., Sprengel, R., Schaefer, A.T. and Fukunaga, I. (2015). Divergent innervation of the olfactory bulb by distinct raphe nuclei. J. Comp. Neurol. 523, 805-813
Tang, W., Szokol, K., Jensen, V., Enger, R., Trivedi, C.A., Hvalby, O., Helm, P.J., Looger, L.L., Sprengel, R. and Nagelhus, E.A. (2015). Stimulation-evoked Ca2+ signals in astrocytic processes at hippocampal CA3-CA1 synapses of adult mice are modulated by glutamate and ATP. J. Neurosci. 35, 3016-3021
Weber, T., Vogt, MA, Gartside, S.E., Berger, S.M., Lujan, R., Lau, T., Herrmann, E., Sprengel, R., Bartsch, D. and Gass, P. (2015). Adult AMPA GLUA1 receptor subunit loss in 5-HT neurons results in a specific anxiety-phenotype with evidence for dysregulation of 5-HT neuronal activity. Neuropsychopharmacology 40, 1471-1484