K. Domenica Karavitaki

Sliding Adhesion Confers Coherent Motion to Hair Cell Stereocilia and Parallel Gating to Transduction Channels

When the tip of a hair bundle is deflected by a sensory stimulus, the stereocilia pivot as a unit, producing a shearing displacement between adjacent tips. It is not clear how stereocilia can stick together laterally but still shear. We used dissociated hair cells from the bullfrog saccule and high-speed video imaging to characterize this sliding adhesion. Movement of individual stereocilia was proportional to height, indicating that stereocilia pivot at their basal insertion points. All stereocilia moved by approximately the same angular deflection, and the same motion was observed at 1, 20, and 700 Hz stimulus frequency. Motions were consistent with a geometric model that assumes the stiffness of lateral links holding stereocilia together is >1000 times the pivot stiffness of stereocilia and that these links can slide in the plane of the membrane—in essence, that stereocilia shear without separation. The same motion was observed when bundles were moved perpendicular to the tip links, or when tip links, ankle links, and shaft connectors were cut, ruling out these links as the basis for sliding adhesion. Stereocilia rootlets are angled toward the center of the bundle, tending to push stereocilia tips together for small deflections. However, stereocilia remained cohesive for deflections of up to ±35°, ruling out rootlet prestressing as the basis for sliding adhesion. These observations suggest that horizontal top connectors mediate a sliding adhesion. They also indicate that all transduction channels of a hair cell are mechanically in parallel, an arrangement that may enhance amplification in the inner ear.

XIRP2, an Actin-Binding Protein Essential for Inner Ear Hair-Cell Stereocilia

Hair cells of the inner ear are mechanoreceptors for hearing and balance, and proteins highly enriched in hair cells may have specific roles in the development and maintenance of the mechanotransduction apparatus. We identified XIRP2/mXinβ as an enriched protein likely to be essential for hair cells. We found that different isoforms of this protein are expressed and differentially located: short splice forms (also called XEPLIN) are targeted more to stereocilia, whereas two long isoforms containing a XIN-repeat domain are in both stereocilia and cuticular plates. Mice lacking the Xirp2 gene developed normal stereocilia bundles, but these degenerated with time: stereocilia were lost and long membranous protrusions emanated from the nearby apical surfaces. At an ultrastructural level, the paracrystalline actin filaments became disorganized. XIRP2 is apparently involved in the maintenance of actin structures in stereocilia and cuticular plates of hair cells, and perhaps in other organs where it is expressed.

Engineering biomimetic hair bundle sensors for underwater sensing applications

We present the fabrication of an artificial MEMS hair bundle sensor designed to approximate the structural and functional principles of the flow-sensing bundles found in fish neuromast hair cells. The sensor consists of micro-pillars of graded height connected with piezoelectric nanofiber “tip-links” and encapsulated by a hydrogel cupula-like structure. Fluid drag force actuates the hydrogel cupula and deflects the micro-pillar bundle, stretching the nanofibers and generating electric charges. These biomimetic sensors achieve an ultrahigh sensitivity of 0.286 mV/(mm/s) and an extremely low threshold detection limit of 8.24 µm/s. A complete version of this paper has been published [1].We present the fabrication of an artificial MEMS hair bundle sensor designed to approximate the structural and functional principles of the flow-sensing bundles found in fish neuromast hair cells. The sensor consists of micro-pillars of graded height connected with piezoelectric nanofiber “tip-links” and encapsulated by a hydrogel cupula-like structure. Fluid drag force actuates the hydrogel cupula and deflects the micro-pillar bundle, stretching the nanofibers and generating electric charges. These biomimetic sensors achieve an ultrahigh sensitivity of 0.286 mV/(mm/s) and an extremely low threshold detection limit of 8.24 µm/s. A complete version of this paper has been published [1].

Design of a Tunable PDMS Force Delivery and Sensing Probe for Studying Mechanosensation

This paper presents the design and analysis of an innovative PDMS probe for investigating mechanotransduction and force generation by mechanosensory cells and organs. The low Youngs modulus of PDMS together with a novel ring-spring probe design allow an easily tunable stiffness of 1-100 mN/m or larger. The probe design limits the fluid drag and allows settling times of 200 μs or less depending on the compliance of the probe. Moreover, the custom-tailored tip geometry of this device allows good coupling to a variety of sensory structures. The probe can be used as a force sensor or a force actuator using force-displacement calibration curves computed by the finite-element method (FEM). Finally, a FEM modal analysis for computing resonant frequencies confirmed that probe resonances are in the high kilohertz range, allowing use across the frequency range of most biological sensors.

PDMS ring-spring soft probe for nano-force biosensing

We present an innovative design for a flexible probe to study mechanisms of biological force sensing and force generation in the piconewton to micronewton range. Made of polydimethylsiloxane (PDMS) and employing a novel ring-spring section with adjustable size, the device works both as a force sensor and force actuator by precise calibration of its tunable stiffness and optical measurement of ring deformation. In addition, the tip geometry of the probe can be properly shaped to fit the anatomical profile of the sensory receptor of interest and to reproduce the in vivo stimulation. Finally, use of Finite Element Method (FEM) modal analysis confirms that the resonance frequencies of probes are outside the frequency range of interest for many sensory systems.

In search of the cochlear amplifier: New mechanical and molecular tools to probe transduction channel function

The study of mechanotransduction in cochlear hair cells requires stimulus methods that mimic the in-vivo stimulation. We have developed a new mechanical probe to better mimic the physiological stimulus delivered to cochlear hair cells through the overlying tectorial membrane. We combine these new probes with electroporation to study the contribution of different components of the transduction apparatus.

Hair Cells: Bundles, Tuning, Transduction—A Moderated Discussion

A discussion moderated by the authors on the topic “Hair Cells: Bundles, Tuning, Transduction” was held on 17 July 2011 at the 11th International Mechanics of Hearing Workshop in Williamstown, Massachusetts. The paper provides an edited transcript of the session.

Hair cells in motion: Imaging the organ of Corti

The mammalian cochlea contains two types of sensory cells, inner hair cells (IHCs) and outer hair cells (OHCs). The IHCs provide the vast majority of the synaptic input to the auditory nerve while the OHCs express a unique motor protein, prestin, and appear to participate in an electromechanical feedback loop that amplifies the motion of the organ of Corti (OC). To study this amplification process we have employed stroboscopic video microscopy to quantify the motion of various elements of the OC. Extracellular electrical stimulation was used to excite OHC motility and a computer‐controlled high‐intensity light‐emitting diode (LED) is used to illuminate the organ OC in an excised cochlear preparation. Motion is measured by extracting small regions of interest (ROIs) from the images and cross‐correlating the ROIs taken during electrical stimulation with a reference image from the same ROIs taken with no stimulation. The observed motion is quite complex with several vibration modes observed. One of the major...