Browsing by Author "Spoon, Corrie E."

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    Biomechanics of hair cell kinocilia: experimental measurement of kinocilium shaft stiffness and base rotational stiffness with Euler-Bernoulli and Timoshenko beam analysis
    Spoon, Corrie E.; Grant, John Wallace (Company of Biologists, 2011-03-01)
    Vestibular hair cell bundles in the inner ear contain a single kinocilium composed of a 9+2 microtubule structure. Kinocilia play a crucial role in transmitting movement of the overlying mass, otoconial membrane or cupula to the mechanotransducing portion of the hair cell bundle. Little is known regarding the mechanical deformation properties of the kinocilium. Using a force-deflection technique, we measured two important mechanical properties of kinocilia in the utricle of a turtle, Trachemys (Pseudemys) scripta elegans. First, we measured the stiffness of kinocilia with different heights. These kinocilia were assumed to be homogenous cylindrical rods and were modeled as both isotropic Euler-Bernoulli beams and transversely isotropic Timoshenko beams. Two mechanical properties of the kinocilia were derived from the beam analysis: flexural rigidity (El) and shear rigidity (kGA). The Timoshenko model produced a better fit to the experimental data, predicting El=10,400 pN mu m(2) and kGA=247 pN. Assuming a homogenous rod, the shear modulus (G=1.9 kPa) was four orders of magnitude less than Young's modulus (E=14.1 MPa), indicating that significant shear deformation occurs within deflected kinocilia. When analyzed as an Euler-Bernoulli beam, which neglects translational shear, El increased linearly with kinocilium height, giving underestimates of El for shorter kinocilia. Second, we measured the rotational stiffness of the kinocilium insertion (kappa) into the hair cell's apical surface. Following BAPTA treatment to break the kinocilial links, the kinocilia remained upright, and kappa was measured as 177 +/- 47 pN mu m rad(-1). The mechanical parameters we quantified are important for understanding how forces arising from head movement are transduced and encoded by hair cells.
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    Hair Bundle Stiffness in the Turtle Utricle: Structural and Regional Variations
    Spoon, Corrie E. (Virginia Tech, 2007-11-19)
    Vestibular hair cells are mechanotransducing sensory receptors in the vertebrate inner ear that detect movement and orientation of the head with respect to gravity. The morphologies of their ciliary bundles vary greatly for different species, endorgans, and within the same endorgan. Bundle morphology in the turtle utricle, like other species, demonstrates highly organized regional variations. These structural differences in bundles impact their mechanical behavior and the process of mechanotransduction. To further understanding of the mechanical behavior of hair bundles, this work experimentally measured the stiffness of bundles with differing morphology, the stiffness contribution of interciliary links and the mechanical properties of the kinocilium in the turtle utricle. The stiffness of hair bundles of varying structure and location along a medial to lateral transect of the utricle was examined. Bundle stiffness was greatest in the striola and demonstrated a systematic decline with location from the line of polarity reversal. The average stiffness of bundles in the striola and extrastriola were 82 ± 46 (n=48) and 9 ± 5 (n=25) µN/m, respectively. The stiff and weak bundles demonstrated characteristic morphologies. The stiffest bundles have short kinocilium, tall stereocilia, and ratios of kinocilium to tallest stereocilia height (KS) close to 1. In contrast, the compliant bundles have tall kinocilium, short stereocilia, and KS ratios ranging from 1.6 – 8. The stiffer bundles also tend to have longer array lengths and steeper slopes. Measurements of bundle stiffness in the turtle utricle are lower than those previously reported which may be attributed to morphological differences between species. The stiffness contributions of the interciliary links were also examined through their selective removal with exposure to the Ca²⁺ chelator BAPTA and the protease subtilisin. BAPTA treatment reportedly breaks tip, kinocilial and ankle links while subtilisin breaks the shaft and ankle links. Following BAPTA and subtilisin treatments, bundle stiffness reduced by 65 ± 10% and 63 ± 11%, respectively. The mechanical properties of the kinocilium were measured with novel techniques. Flexural rigidity (EI) was measure while the kinocilium was fixed at the height of the tallest stereocilia using a glass supporting probe. Through both force deflection and a high speed video technique, measured values of EI ranged from 1460 – 6150 pN·µm2. The rotational stiffness of the kinocilium about its apical insertion was also measured. Bundles were treated with BAPTA to break the kinocilial links and separate the kinocilium from neighboring stereocilia. Using a force deflection technique, the rotational stiffness of the kinocilium was measured as 120 ± 17 pN·µm/rad.
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    Somersault of Paramecium in extremely confined environments
    Jana, Saikat; Eddins, Aja; Spoon, Corrie E.; Jung, Sunghwan (Springer Nature, 2015-08-19)
    We investigate various swimming modes of Paramecium in geometric confinements and a non-swimming self-bending behavior like a somersault, which is quite different from the previously reported behaviors. We observe that Paramecia execute directional sinusoidal trajectories in thick fluid films, whereas Paramecia meander around a localized region and execute frequent turns due to collisions with adjacent walls in thin fluid films. When Paramecia are further constrained in rectangular channels narrower than the length of the cell body, a fraction of meandering Paramecia buckle their body by pushing on the channel walls. The bucking (self-bending) of the cell body allows the Paramecium to reorient its anterior end and explore a completely new direction in extremely confined spaces. Using force deflection method, we quantify the Young's modulus of the cell and estimate the swimming and bending powers exerted by Paramecium. The analysis shows that Paramecia can utilize a fraction of its swimming power to execute the self-bending maneuver within the confined channel and no extra power may be required for this new kind of self-bending behavior. This investigation sheds light on how micro-organisms can use the flexibility of the body to actively navigate within confined spaces.