Involvement of TRPV4 channels in Aβ(40)-induced hippocampal cell death and astrocytic Ca(2+) signalling.
Previous studies suggested that amyloid β (Aβ)-induced disruption of astrocytic Ca(2+) signalling and oxidative stress play a major role in the progression towards neuronal and glial death in Alzheimer's disease. We have recently demonstrated that Ca(2+)-permeable TRPV4 channels are highly expressed in rat hippocampal astrocytes and are involved in oxidative stress-induced cell damage. The aim of this study was to test the hypothesis that TRPV4 channels also contribute to hippocampal damage evoked by Aβ. Synthetic Aβ40 evoked cell death in hippocampal slice cultures in a concentration (0-20μM) and time (12-48h) dependent manner, after cultures were preconditioned with sublethal concentration of buthionine sulfoximine (1.5μM) which enhanced endogenous ROS production. As demonstrated by propidium iodide fluorescence, damage was observed in the granule cell layer of the dentate gyrus and to a smaller degree in pyramidal neurons of the CA1-CA3 region, as well as in glia cells mainly at the edge of the slice. Immunocytochemistry revealed an altered pattern of TRPV4 and GFAP protein expression, and reactive astrogliosis surrounding pyramidal CA1-CA3 neurons. Neuronal and astrocytic damage was attenuated by the antioxidant Trolox, TRPV4 channel blockers Gd(3+) and ruthenium red (RR), and a specific inhibitor of the redox and Ca(2+)-sensitive phospholipase A2 enzyme (MAFP). In disassociated co-cultures of hippocampal neurons and astrocytes without BSO preconditioning, Aβ40 evoked pronounced neuronal damage, enhanced the expression of TRPV4 and GFAP proteins (indicative of reactive astrogliosis), and increased intracellular free Ca(2+) concentration in astrocytes. The latter effect was attenuated by RR and in Ca(2+)-free media. These data show that Aβ40 can activate astrocytic TRPV4 channels in the hippocampus, leading to neuronal and astrocytic damage in a Ca(2+) and oxidative stress-dependent manner.
Department of Physiology, Faculty of Medical and Health Sciences, University of Auckland, Private Bag 92-019, Auckland, New Zealand. Electronic address: j.bai@auckland.ac.nz.
Ji-Zhong
JZ
Bai
Department of Physiology, Faculty of Medical and Health Sciences, University of Auckland, Private Bag 92-019, Auckland, New Zealand.
Janusz
J
Lipski
D016229:Amyloid beta-Peptides / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D000975:Antioxidants / Q000494:pharmacology; D001095:Arachidonic Acids / Q000494:pharmacology; D001253:Astrocytes / Q000187:drug effects*; D019256:Cadmium Chloride / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects*; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology; D018920:Coculture Techniques; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology*; D009474:Neurons / Q000187:drug effects*; D009924:Organ Culture Techniques; D063065:Organophosphonates / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D050916:TRPV Cation Channels / Q000378:metabolism*; D013997:Time Factors
D016229:Amyloid beta-Peptides; D000975:Antioxidants; D001095:Arachidonic Acids; D002839:Chromans; D004791:Enzyme Inhibitors; D063065:Organophosphonates; D010446:Peptide Fragments; D050916:TRPV Cation Channels; C486577:Trpv4 protein, rat; C078592:amyloid beta-protein (1-40); C104810:methyl arachidonylfluorophosphonate; D019256:Cadmium Chloride; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid
Amyloid ß-peptide; Buthionine sulfoximine; Ca(2+)signalling; Disassociated hippocampal culture; Organotypic hippocampal culture; Oxidative stress