Supplementary Materialsawz288_Supplementary_Data. analysis. Specifically, we discovered that A2AR overexpression in THY-Tau22 mice resulted in the hippocampal upregulation of C1q go with proteinalso seen in individuals with frontotemporal lobar degenerationand correlated with the increased loss of glutamatergic synapses, most likely underlying the noticed memory space deficits. These data reveal an integral effect of overactive neuronal A2AR in the starting point of synaptic reduction in tauopathies, paving the true method for new therapeutic approaches. gene coding tau (Hutton P301L mutation. Promoting neuronal A2AR upregulation inside a tauopathy mouse model (THY-Tau22) resulted in a hippocampal upregulation of C1q go with protein connected with a lack of glutamatergic synapses and a potentiation of spatial memory space deficits, suggesting an instrumental role of neuronal A2AR dysregulation towards tau pathology-induced cognitive alterations. Materials and methods Post-mortem brain samples Post-mortem brain tissue was obtained from brain banks at university medical centres in Lille (France), Paris (France) and Geneva (Switzerland), following approval by the local institutional review board and the provision of written, informed consent by the donors family. We used samples from the temporal cortex of three FTLD-tau patients with P301L mutation (Forrest 0.5, Students > 0.5, ANOVA; mean SEM). They did not differ in the mean post-mortem interval (FTLD-tau P301L, 18.0 9.8 h; control group A, 22.0 7.5 h; 0.78; Students > 0.5, ANOVA; mean SEM). Most participants and methods have been described previously (Huin gene, the bidirectional inducible access to food (SafeA04) and water. The animals were maintained in compliance with European standards for the care and use of laboratory animals and experimental protocols approved by the local Animal Ethical Committee (agreement #12787-2015101320441671 v9 from CEEA75, Lille, France). The overexpression of mouse A2AR in forebrain neurons was achieved by crossing the in-house developed TRE-A2AR transgenic RSV604 R enantiomer strain (in which mouse receptor cDNA is under the control of a Tet-responsive element) and the transgenic CaMKII-tTA line, expressing the tetracycline-controlled transactivator protein (tTA) under regulatory control of the forebrain-specific calcium-calmodulin-dependent kinase II (CaMKII) promoter [B6.Cg-Tg(Camk2a-tTA)1Mmay/DboJ; SN 7004; The Jackson Laboratory; Fig. 2A]. Previously, the tTA transactivator was found to favour hippocampal atrophy in non-C57Bl6 genetic backgrounds (Han panels represent immunostainings at the level of the striatum and panels at the level of the hippocampus and cortex. Scale bar = 1 mm. (D) Co-immunostainings with A2AR (red) and either neuronal (NeuN), microglial (Iba1) or astrocytic (GFAP and S100) markers (green) showing the neuronal-specificity of A2AR overexpression in CaMKII-tTA/TRE-A2AR mice. DAPI (blue) represents cell nuclei. Scale bar = 20 m. (E) Co-immunostainings between A2AR (red), NeuN (as marker of RSV604 R enantiomer mature neurons, white) Rabbit polyclonal to HERC4 and doublecortin (DCX, as marker of immature neurons, green) in CaMKII-tTA/TRE-A2AR mice (A2AR). A2AR was not expressed in immature neurons. Scale bar = 100 m. (F) Averaged time course of field excitatory postsynaptic potentials (fEPSP) after perfusion with “type”:”entrez-protein”,”attrs”:”text”:”SCH58261″,”term_id”:”1052882304″,”term_text”:”SCH58261″SCH58261 (50 nM) for 30 min on hippocampal slices from wild-type and double CaMKII-tTA/TRE-A2AR transgenic mice (*0.05, 5 per group). A2AR blockade significantly inhibited fEPSPs in double transgenic mice suggesting a gain of function of A2AR upon their overexpression, whereby A2AR exerts a tonic control on basal synaptic transmission, a phenomenon that is not observed in wild-type animals. Generation of RSV604 R enantiomer a new transgenic model of forebrain A2AR overexpression in a THY-Tau22 background THY-Tau22 mice (C57BL6/J background; Schindowski and and directions and between 4 m and 6 m in depth of the stack (4/group) were generated from 300 ng of total RNA using Illumina TruSeq RNA Sample Preparation Kit v2 (Illimina RS-122-2101). Briefly,.