Hair cells of the inner ear exhibit an active process, believed to be crucial for achieving the sensitivity of auditory and vestibular detection. One of the manifestations of the active process is the occurrence of spontaneous hair bundle oscillations in vitro. Hair bundles are coupled by overlying membranes in vivo; hence, explaining the potential role of innate bundle motility in the generation of otoacoustic emissions requires an understanding of the effects of coupling on the active bundle dynamics. We used microbeads to connect small groups of hair cell bundles, using in vitro preparations that maintain their innate oscillations. Our experiments demonstrate robust synchronization of spontaneous oscillations, with either 1:1 or multi-mode phase-locking. The frequency of synchronized oscillation was found to be near the mean of the innate frequencies of individual bundles. Coupling also led to an improved regularity of entrained oscillations, demonstrated by an increase in the quality factor.
Mechanosensation is the process by which cells sense mechanical forces and translate them into electrical and chemical signals. Important physiological functions including sensation of touch, sensation of sound and proprioception are based on mechanosensation. Recent studies identified the molecular identity of a highly conserved group of mechanosensitive receptors, called Piezo receptors, which are both necessary and sufficient for cells to sense force. Currently, structure-function information of Piezo receptors is limited, and the mechanism how these receptors sense force, which conformational changes they undergo, and which precise mechanical stimuli cause these channels to open, remains poorly understood. Studies to date have used rather brute forces reaching several hundrets of nano-Newton to mechanically activate Piezo channels, which are likely too high to be of specific nature. To manipulate Piezo receptors at high precision and accuracy in a cellular context, new techniques have to be applied. We are currently developing assays to investigate Piezo receptors at the molecular level with delicate force control. Using a combination of high-resolution atomic force microscopy (AFM) and time-lapse fluorescent calcium imaging, we aim to find more effi
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