caa a22 4500 606177027 CHVBK 20210128100750.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00018-015-1949-4 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1949-4 Polymeric alkylpyridinium salts permit intracellular delivery of human Tau in rat hippocampal neurons: requirement of Tau phosphorylation for functional deficits [Elektronische Daten] [Dave Koss, Lianne Robinson, Anna Mietelska-Porowska, Anna Gasiorowska, Kristina Sepčić, Tom Turk, Marcel Jaspars, Grazyna Niewiadomska, Roderick Scott, Bettina Platt, Gernot Riedel] Patients suffering from tauopathies including frontotemporal dementia (FTD) and Alzheimer's disease (AD) present with intra-neuronal aggregation of microtubule-associated protein Tau. During the disease process, Tau undergoes excessive phosphorylation, dissociates from microtubules and aggregates into insoluble neurofibrillary tangles (NFTs), accumulating in the soma. While many aspects of the disease pathology have been replicated in transgenic mouse models, a region-specific non-transgenic expression model is missing. Complementing existing models, we here report a novel region-specific approach to modelling Tau pathology. Local co-administration of the pore-former polymeric 1,3-alkylpyridinium salts (Poly-APS) extracted from marine sponges, and synthetic full-length 4R recombinant human Tau (hTau) was performed in vitro and in vivo. At low doses, Poly-APS was non-toxic and cultured cells exposed to Poly-APS (0.5µg/ml) and hTau (1µg/ml;~22µM) had normal input resistance, resting-state membrane potentials and Ca2+ transients induced either by glutamate or KCl, as did cells exposed to a low concentration of the phosphatase inhibitor Okadaic acid (OA; 1nM, 24h). Combined hTau loading and phosphatase inhibition resulted in a collapse of the membrane potential, suppressed excitation and diminished glutamate and KCl-stimulated Ca2+ transients. Stereotaxic infusions of Poly-APS (0.005µg/ml) and hTau (1µg/ml) bilaterally into the dorsal hippocampus at multiple sites resulted in hTau loading of neurons in rats. A separate cohort received an additional 7-day minipump infusion of OA (1.2nM) intrahippocampally. When tested 2weeks after surgery, rats treated with Poly-APS+hTau+OA presented with subtle learning deficits, but were also impaired in cognitive flexibility and recall. Hippocampal plasticity recorded from slices ex vivo was diminished in Poly-APS+hTau+OA subjects, but not in other treatment groups. Histological sections confirmed the intracellular accumulation of hTau in CA1 pyramidal cells and along their processes; phosphorylated Tau was present only within somata. This study demonstrates that cognitive, physiological and pathological symptoms reminiscent of tauopathies can be induced following non-mutant hTau delivery into CA1 in rats, but functional consequences hinge on increased Tau phosphorylation. Collectively, these data validate a novel model of locally infused recombinant hTau protein as an inducer of Tau pathology in the hippocampus of normal rats; future studies will provide insights into the pathological spread and maturation of Tau pathology. Springer Basel, 2015 Alzheimer's disease nationallicence Frontotemporal dementia nationallicence Tauopathies nationallicence Phospho-Tau nationallicence Poly-APS nationallicence Tau nationallicence LTP nationallicence Calcium nationallicence Koss Dave School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut Robinson Lianne School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut Mietelska-Porowska Anna Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland aut Gasiorowska Anna Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland aut Sepčić Kristina Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia aut Turk Tom Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia aut Jaspars Marcel Department of Chemistry, Marine Biodiscovery Centre, University of Aberdeen, Aberdeen, UK aut Niewiadomska Grazyna Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland aut Scott Roderick School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut Platt Bettina School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut Riedel Gernot School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut Cellular and Molecular Life Sciences Springer International Publishing 72/23(2015-12-01), 4613-4632 1420-682X 72:23<4613 2015 72 18 https://doi.org/10.1007/s00018-015-1949-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1949-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Koss Dave School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- Robinson Lianne School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- Mietelska-Porowska Anna Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland aut NATIONALLICENCE 700 1- Gasiorowska Anna Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland aut NATIONALLICENCE 700 1- Sepčić Kristina Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia aut NATIONALLICENCE 700 1- Turk Tom Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia aut NATIONALLICENCE 700 1- Jaspars Marcel Department of Chemistry, Marine Biodiscovery Centre, University of Aberdeen, Aberdeen, UK aut NATIONALLICENCE 700 1- Niewiadomska Grazyna Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland aut NATIONALLICENCE 700 1- Scott Roderick School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- Platt Bettina School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 700 1- Riedel Gernot School of Medical Sciences, University of Aberdeen, Foresterhill, AB25 2ZD, Aberdeen, UK aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer International Publishing 72/23(2015-12-01), 4613-4632 1420-682X 72:23<4613 2015 72 18