Biology subjects

Ishibashi, Y.

Publications and source records attributed to Ishibashi, Y..

2 recordsLinked to original sources

bioRxiv2024

Live-cell single-molecule fluorescence microscopy for protruding organelles reveals regulatory mechanisms of MYO7A-driven cargo transport in stereocilia of inner ear hair cells

Stereocilia are unidirectional F-actin-based cylindrical protrusions on the apical surface of inner ear hair cells and function as biological mechanosensors of sound and acceleration. Development of functional stereocilia requires motor activities of unconventional myosins to transport proteins necessary for elongating the F-actin cores and to assemble the mechanoelectrical transduction (MET) channel complex. However, how each myosin localizes in stereocilia using the energy from ATP hydrolysis is only partially understood. In this study, we develop a methodology for live-cell single-molecule fluorescence microscopy of organelles protruding from the apical surface using a dual-view light-sheet microscope, diSPIM. We demonstrate that MYO7A, a component of the MET machinery, traffics as a dimer in stereocilia. Movements of MYO7A are restricted when scaffolded by the plasma membrane and F-actin as mediated by MYO7As interacting partners. Here, we discuss the technical details of our methodology and its future applications including analyses of cargo transportation in various organelles.

cell biology↗

bioRxiv2022

Large-area electrical imaging having single neuron resolution using 236,880 electrodes CMOS-MEA technology

The electrophysiological technology having a high spatio-temporal resolution at the single-cell level, and noninvasive measurements of large areas provides insights on underlying neuronal function. Here, we used a complementary metal-oxide semiconductor (CMOS)-microelectrode array (MEA) that uses 236,880 electrodes each with an electrode size of 11.22 x 11.22 {micro}m and 236,880 covering a wide area of 5.5 x 5.7 mm in presenting a detailed and single-cell-level neural activity analysis platform for brain slices, human iPS cell-derived cortical networks, peripheral neurons, and human brain organoids. Propagation pattern characteristics between brain regions changes the synaptic strength into compounds based on single-cell time-series patterns, classification based on single DRG neuron firing patterns and compound responses, axonal conduction characteristics and changes to anticancer drugs, and network activities and transition to compounds in brain organoids were extracted. This detailed analysis of neural activity at the single-cell level using our CMOS-MEA provides a new understanding the basic mechanisms of brain circuits in vitro and ex vivo, on human neurological diseases for drug discovery, and compound toxicity assessment.

neuroscience↗