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Shimamura, R.

Publications and source records attributed to Shimamura, R..

2 recordsLinked to original sources

Multi-modal ion-transport function of potassium channelrhodopsins

Light-gated K+ channelrhodopsins (KCRs) are recognized as an ideal neuronal silencing tool in optogenetics. KCRs contain functionally important residues for a light-driven H+ pump bacteriorhodopsin, but their roles have been unclear. Utilizing electrophysiological methods, we demonstrate that KCRs function as a light-driven H+ pump in the absence of K+. Transport experiments using pH electrodes further elucidated the function of the H+-pump, which operates even in the presence of K+. The hypothesis that the H+ pump exerts a blocking effect on K+ transport in KCRs has been postulated. Furthermore, we ascertained that there was conversion of KCRs from the H+ pump into the H+ channel by light, yet this conversion was never reversed. However, the addition of K+ resulted in the formation of the K+ channel. KCRs exhibit three distinct functional states, H+ pump, H+ channel, and K+ channel. These multifaceted functions are modulated by environmental ions and light.

biophysics↗

Highly efficient induction of functionally mature excitatory neurons from feeder-free human ES/iPS cells

Cortical excitatory neurons (Cx neurons) are the most dominant neuronal cell type in the cerebral cortex, which play a central role in cognition, perception, intellectual behavior and emotional processing. Robust in vitro induction of Cx neurons may facilitate as a tool for the elucidation of brain development and pathomechanism of the intractable neurodevelopmental and neurodegenerative disorders including Alzheimers disease, and thus potentially contribute to drug development. Here, we report a defined method for efficient induction of Cx neurons from the feeder-free-conditioned human embryonic stem cells (ES cells) and induced pluripotent stem cells (iPS cells). By using this method, human ES/iPS cells could be differentiated into ~99% MAP2-positive neurons by three weeks, and these induced neurons, within five weeks, presented various characteristics of mature excitatory neurons such as strong expression of glutamatergic neuron-specific markers (subunits of AMPA and NDMA receptors and CAMKII), highly synchronized spontaneous firing and excitatory postsynaptic current (EPSC). Moreover, the Cx neurons showed susceptibility to the toxicity of A{beta}42 oligomers and excitotoxicity of excessive glutamates, which is another advantage in terms of toxicity test and searching for the therapeutic agents. Taken together, this study provides a novel research platform for the study of neural development and degeneration based on the feeder-free human ES/iPS cell system.

neuroscience↗