Temperature dependency of cupular mechanics and hair cell frequency selectivity in the fish canal lateral line organ

The mechanical frequency selectivity of the cupula located in the supraorbital lateral line canal and the frequency selectivity of the hair cells driven by the cupula were measured simultaneously in vivo. Laser interferometry was used to measure cupular mechanics and extracellular receptor potentials were recorded to determine hair cell frequency selectivity. Results were obtained from two teleost fish species, the ruffe (Acerina cernua L.), a European temperate zone freshwater fish, and the tropical African knife fish (Xenomiystus nigri). In both species cupular displacement grows with increasing frequency of canal fluid displacement, reaching a maximum at 115 Hz in the ruffe and at 460 Hz in the African knife fish. Cupular best frequencies were independent of temperature. Cut-off frequencies of hair cell frequency selectivity were found to depend on temperature with a Q10 of 1.75, ranging from 116 Hz (4 degrees C) to 290 Hz (20 degrees C), as established in the ruffe. At normal habitat temperatures of the two fish species (ruffe, 4 degrees C; African knife fish, 28 degrees C), this results in hair cell cut-off frequencies that match the two different cupular best frequencies remarkably well. This match suggests adjusted signal transfer in these two peripheral stages of canal lateral line transduction.     

Rapid responses of the cupula in the lateral line of ruffe (<Emphasis Type="Italic">Gymnocephalus cernuus</Emphasis>)

Displacements of cupulae in the supraorbital lateral line canal in ruffe (Gymnocephalus cernuus) have been measured using laser interferometry and by applying transient as well as sinusoidal fluid stimuli in the lateral line canal. The cupular displacement in response to impulses of fluid velocity shows damped oscillations at approximately 120 Hz and a relaxation time-constant of 4.4 ms, commensurate with a quality factor of approximately 1.8. These values are in close agreement with the frequency characteristics determined via sinusoidal fluid stimuli, implying that the nonlinearity of cupular dynamics imposed by the gating apparatus of the sensory hair cells is limited in the range of cupular displacements and velocities measured (100–300 nm; 100–300 m/s). The measurements also show that cupular displacement instantaneously follows the initial waveform of transient stimuli. The functional significance of the observed cupular dynamics is discussed.     

Mechanical filtering by the boundary layer and fluid–structure interaction in the superficial neuromast of the fish lateral line system

A great diversity of aquatic animals detects water flow with ciliated mechanoreceptors on the body's surface. In order to understand how these receptors mechanically filter signals, we developed a theoretical model of the superficial neuromast in the fish lateral line system. The cupula of the neuromast was modeled as a cylindrical beam that deflects in response to an oscillating flow field. Its accuracy was verified by comparison with prior measurements of cupular deflection in larval zebrafish (Danio rerio). The model predicts that the boundary layer of flow over the body attenuates low-frequency stimuli. The fluid-structure interaction between this flow and the cupula attenuates high-frequency stimuli. The number and height of hair cell kinocilia and the dimensions of the cupular matrix determine the range of intermediate frequencies to which a neuromast is sensitive. By articulating the individual mechanical contributions of the boundary layer and the components of cupular morphology, this model provides the theoretical framework for understanding how a hydrodynamic receptor filters flow signals.     

A mathematical model for top-shelf vertigo: the role of sedimenting otoconia in BPPV

Benign paroxysmal positional vertigo (BPPV) is a mechanical disorder of the vestibular system in which calcite particles called otoconia interfere with the mechanical functioning of the fluid-filled semicircular canals normally used to sense rotation. Using hydrodynamic models, we examine the two mechanisms proposed by the medical community for BPPV: cupulolithiasis, in which otoconia attach directly to the cupula (a sensory membrane), and canalithiasis, in which otoconia settle through the canals and exert a fluid pressure across the cupula. We utilize known hydrodynamic calculations and make reasonable geometric and physical approximations to derive an expression for the transcupular pressure DeltaPc exerted by a settling solid particle in canalithiasis. By tracking settling otoconia in a two-dimensional model geometry, the cupular volume displacement and associated eye response (nystagmus) can be calculated quantitatively. Several important features emerge: (1) a pressure amplification occurs as otoconia enter a narrowing duct; (2) an average-sized otoconium requires approximately 5 s to settle through the wide ampulla, where DeltaPc is not amplified, which suggests a mechanism for the observed latency of BPPV; and (3) an average-sized otoconium beginning below the center of the cupula can cause a volumetric cupular displacement on the order of 30 pL, with nystagmus of order 2 degrees/s, which is approximately the threshold for sensation. Larger cupular volume displacement and nystagmus could result from larger and/or multiple otoconia.     

Cupular organs in two species of Corella (Tunicata: Ascidiacea)

Abstract. Simple cupular organs similar to those described in Ciona intestinalis were observed in Corella eumyota. They consist of a macula containing the cell bodies of 20–30 primary sensory neurons whose cilia project into a dome‐ or finger‐shaped structure, the cupula. Rather than being found in the mantle lining as in C. intestinalis, the organs were located on the atrial surface of the branchial sac. The sensory innervation was examined in whole‐mount preparations using anti‐tubulin immunohistochemistry. Sensory neurons in C. eumyota showed no immunoreactivity with antisera raised against gonadotropin‐releasing hormone (GnRH). A novel, elongated sense organ termed the cupular strand was found in Corella inflata. It has the same basic components as the simple type of cupular organ but consists of a single, long structure containing ～1500 sensory cells. Located on the atrial surface of the branchial sac, it extends along the midline of the dorsal fold, from the gonoduct openings almost as far as the brain. Preparations were examined using optical and electron microscopy. Nerves and cilia were visualized by anti‐tubulin immunofluorescence microscopy. It was possible to follow the sensory axons from the macula of the cupular strand to points where they joined branches of the visceral nerve, which enters a nerve root at the back of the brain. In C. inflata the sensory cell bodies and their axons were immunoreactive not only with anti‐tubulin but also with an antiserum raised against Tunicate I GnRH. There was no immunoreactivity, however, with Chicken II and catfish GnRH antisera. All three GnRH antisera labeled the dorsal strand plexus, a structure associated with production of GnRH in its role as a reproductive hormone. We concluded that the GnRH‐like molecule labeled in sensory neurons differs from the form of GnRH found in the dorsal strand plexus, and may have a different function, perhaps in the neural control of ciliary activity. The function of the cupular organs in species of Corella has not yet been investigated physiologically, but by analogy with such structures in other metazoans, cupular organs are probably hydrodynamic sensors registering local disturbances or changes in water flow through the atrial cavity.     

Topography and mechanics of the cupula in the fish lateral line. I. Variation of cupular structure and composition in three dimensions.

The cupula of the supraorbital neuromast in the lateral line canal of the clown knifefish contains vertical columns. In the central region of the cupula overlying the macula, these columns are densely packed, are relatively constant in size, and run from the base of the cupula to the surface of the cupula which is exposed to canal fluid. There are two types of columns, dark and light, which form elliptical compartments in planes of section that cut across the columns; the cupula therefore has the appearance of mosaic tile in such sections. The dark compartments contain tubules that extend from the base of the cupula at the junction with the macula to the top of the cupula. Each tubule is associated with the kinocilium of a single hair cell. The lateral parts of the cupula, not overlying the macula, also contain compartments, but these compartments differ in size and structure from those in the central region. In addition to the compartments, the central region of the cupula also contains spherical aggregates of droplets. These small aggregates, termed mora, are found principally in a layer within the central region of the cupula, but are also found outside this layer. Because of their light-reflecting properties, the mora can be used for noninvasive optical measurements in vivo of the motion of the cupula.     

Ultrastructure of free neuromasts of Bathygobius fuscus (gobiidae) and canal neuromasts of Apogon cyanosoma (apogonidae)

Light and electron microscopic observations were made on the lateral line organs of the free neuromasts of the goby Bathygobius fuscus and the canal neuromasts of the cardinal fish Apogon cyanosoma. As in other lateral line systems, each neuromast consists of hair cells, supporting cells and mantle supporting cells, the whole being covered by a cupula. In B. fuscus the free neuromasts are mounted on papillae and have hair cells with stereocilia up to 2.5 μm long and a single kinocilium at least 25 μm long. Each neuromast is covered by a vane-like cupula that can be divided into two regions. The central region over the sensory area contains columns of myelin-like figures. These figures are absent from the outer region covering the mantle. The canal neuromasts of A. cyanosoma are diamond-shaped with up to 1,500 hair cells. The cupula is unusual in having a channel that lies over the sensory region. The hair cells have up to 45 stereocilia, the tallest reaching 2.5 μm, and a kinocilium at least 5 μm long. Tip links are shown for the first time between rows of stereocilia of the hair cells of lateral line neuromasts. The presence of tip links has now been demonstrated for all acousticolateral hair cell systems.     

A poroelastic continuum model of the cupula partition and the response dynamics of the vestibular semicircular canal.

Using mixture theory, an axisymmetric continuum model is presented describing the response dynamics of the vestibular semicircular canals to canal-centered head rotation in which the cupula partition is modeled as a poroelastic mixture of interpenetrating solid and fluid constituents. The solid matrix of the cupula is assumed to behave as a linear elastic material, whereas the fluid constituent is assumed to be Newtonian. A regular perturbation analysis of the fluid dynamics in the canal provides a dynamic boundary condition, which acts across the cupula partition. Numerical solution of the coupled system of momentum equations provides the spatio-temporal displacement fields for both the fluid and solid constituents of the cupula. Results indicate that at frequencies above 1 Hz, the fluid constituent is dynamically entrained by the solid matrix such that their motions are bound as if to exist as a single component. The resulting high-frequency response is consistent with the macromechanical response predicted by single-component viscoelastic models of the cupula. Below 1 Hz, the dynamic coupling between the fluid and solid constituents weakens and the transcupular differential pressure is sufficient to force fluid through the mixture with little deformation of the solid matrix. Results are sensitive to the precise value of the cupular permeability. One of the most important distinctions between the present analysis and previous impermeable models of the cupula arises at the micromechanical level in terms of the local fluid flow that is predicted to occur within the cupula and around the ciliary bundles and sensory hair cells. Another important result reveals that the permeation dynamics predicted below 1 Hz gives rise to the same low-frequency macromechanical response as would occur with an impermeable viscoelastic structure having a much greater stiffness. Current estimates of the mechanical stiffness of the cupula, based solely on afferent nerve data, may therefore overestimate the true value intrinsic to the solid matrix by as much as an order of magnitude.     

Numerical modeling of the cupular displacement and motion of otoconia particles in a semicircular canal

Balance is achieved and maintained by a balance system called a labyrinth that is composed of three semicircular canals and the otolith organs that sense linear gravity and acceleration. Within each semicircular canal, there is a gelatinous structure called the cupula, which is deformed under the influence of the surrounding endolymph. One of the balance disorders is benign paroxysmal positional vertigo, and one of the pathological conditions that have been identified as possible causes of this syndrome is canalithiasis—disturbance of the endolymph flow and cupular displacement caused by the free-moving otoconia particles within the lumen of the canal. Analysis of phenomena occurring within the semicircular canal can help to explain some balance-related disorders and the response of the vestibular system to external perturbations under various pathological conditions. Numerical simulations allow a study of the influence of a wide range of factors, without the need to perform experiments and clinical examinations. In case of canalithiasis, an accurate explanation and tracking of the motion of otoconia particles in vivo is obviously nearly impossible. In this study, a numerical model was developed to predict the motion of otoconia particles within the semicircular canal and the effect of the endolymph flow and particles on the deformation of the cupula.     

An Electrokinetic Model of Transduction in the Semicircular Canal

Transduction in the semicircular canal was studied by focusing an infrared beam on either side of exposed ampullae from the posterior canals of Rana pipiens. The direction of fluid movement resulting from a stimulus was inferred by observing the polarity of the change in afferent impulse mean rate relative to the spontaneous value. On the basis of the accepted functional polarization of this receptor, the results indicate that fluid moved toward the warmer side of the ampulla. Convection and thermal reception were shown to be unlikely explanations for these results. Morover, cupular displacements toward the warmer side would not be expected. Because thermo-osmosis can cause fluid to move toward the warmer side in a gelatin membrane, the results can be interpreted as evidence that thermo-osmosis occurred in the gelatinous cupula and influenced the transduction mechanism. Thermo-osmosis of liquids appears to be due to an electric field that is set up in a charged membrane; hence, the hair cells might have detected an electric field that occurred in the cupula during thermo-osmosis. Electroreception might be an important link in the transduction of physiological stimuli also. Rotational stimuli could result in weak electric fields in the cupula by the mechanoelectric effect. Cupular displacements could be important for large stimuli, but extrapolations to threshold stimuli suggest displacements of angstrom amplitudes. Therefore, electroreception by the hair cells could be an explanation of the great sensitivity that has been observed in the semicircular canal and other labyrinthine receptors.     

A comparison of Lateral Line Morphology of Blue Cod and Torrentfish: Two Sandperches of the Family Pinguipedidae

Parapercis colias (blue cod) and Cheimarrichthys fosteri (torrentfish) are two members of the family Pinguipedidae. They reside in habitats with different background levels of hydrodynamic activity and differ in their feeding ecology. The peripheral morphology of the mechanosensory lateral line system was investigated in each species. The torrentfish is the only freshwater member of this otherwise exclusively marine family. It resides in turbulent fast flowing habitats and feeds nocturnally on stream drift. Torrentfish have many superficial neuromasts and a simple unbranched canal system. In comparison the blue cod resides in sub-tidal slow flowing habitats, is a diurnal predator and has relatively few superficial neuromasts and a well-developed branching canal system. For these two species the background level of hydrodynamic activity does not appear to be the dominant selection pressure on lateral line morphology, in the case of the torrentfish in particular it is more compelling to view lateral line morphology in the light of environmental pressures that have favoured the evolution of nocturnal feeding.     

Functional physiology of the semicircular canal ampulla.

Physiology of the semicircular canal (sc) was studied by applying different manipulations to the isolated frog sc. Function of the cupula was investigated by mapping out the mechanical sensitivity on the cupular surface and by removing and replacing the cupula. The cupula was found to be most essential for effective activation of sc receptors. Responses of sc receptors to direct temperature change were studied. The sc nerve discharge increased and decreased due to cool and warm temperature change respectively. This suggests a possibility of direct temperature effect as one of the mechanisms of caloric response.     

Lateral line mediated rheotaxis in the Antarctic fish Pagothenia borchgrevinki

The sensory basis of rheotaxis was investigated in Pagothenia borchgrevinki utilising a laminar flow chamber. The threshold for P. borchgrevinki to exhibit an unconditioned rheotactic response lay between 1 and 2 cm s⁻¹. Disabling the entire lateral line or the superficial neuromast receptors increased the rheotactic threshold to greater than 5 cm s⁻¹. Pharmacological blocking of the lateral line canal system alone had no effect. This study provides a direct demonstration that the superficial lateral line system is involved in mediating rheotaxis. These results, coupled with previous work on Antarctic fishes, suggest a division of labour exists between the two submodalities of the lateral line system. Superficial neuromasts are more responsive to unmodulated flows (DC) and mediate behaviour such as rheotaxis, whereas canal neuromasts detect acceleration components of modulated flows (AC) and are more concerned with behaviour such as feeding. Accepted: 27 October 1998     

The sensory basis of rheotaxis in the blind Mexican cave fish, Astyanax fasciatus

The sensory basis of rheotaxis (orientation to currents) was investigated in the blind Mexican cave fish, Astyanax fasciatus. An unconditioned rheotactic response to uniform velocity flows was exhibited, with a threshold of less than 3 cm s⁻¹. Disabling the entire lateral line or the superficial neuromast receptor class increased the rheotactic threshold to greater than 9 cm s⁻¹. A pharmacological block of the lateral line canal system alone had no effect. These results demonstrate that the superficial lateral line system controls rheotaxis at low current velocities. The effect of pairing an odor stimulant with the water current dropped the rheotactic threshold to less than 0.4 cm s⁻¹. This study provides a clear behavioral role for the superficial neuromasts where none previously existed, and also establishes a link between the mechanosensory lateral line and olfactory systems in the olfactory search behavior of the cave fish. Accepted: 9 January 1999     

Physical dimensions influence the functional property of the semicircular canal

The functional dependence of the semicircular canal upon its physical dimensions was evaluated by measuring the internal radius (r), the radius of curvature (R) and the cupula radius (rC) of the posterior canal in 10 freshly dissected frog labyrinths. These values have been compared to the same parameters of the cat labyrinth. The coefficients I, B, K in the Steinhausen equation were determined for both animals. The A ratio between cupula deflection and endolymph displacement was also calculated by utilizing the Bernard equation. The A ratio is three times larger in the frog than in the cat. It follows that if the same acceleration produces similar endolymph displacements in the posterior canal of both animals, the cupula deflection will be larger in the frog. The solution of the Steinhausen equation in the presence of a constant acceleration, however, reveals that the same stimulus intensity will result in a larger endolymph displacement in the cat posterior canal; similarly, the endolymph displacement directly depends on the duration of the stimulating period in both animals. Contrary to the Bernard assumption, these effects generate a G ratio (psi frog/psi cat) which is less than the Q ratio (A frog/A cat). Moreover, G decreases on increasing the duration of the stimulating period. For stimuli of short duration the semicircular canal of a small animal is expected to exhibit a higher sensitivity than that of a larger one. However, the definitive primary afferent discharge will be largely controlled by the receptor/generator potential properties.     

Peripheral and central processing of lateral line information

The lateral line is a hydrodynamic sensory system that allows fishes and aquatic amphibians to detect the water motions caused, for instance, by conspecifics, predators or prey. Typically the peripheral lateral line of fishes consists of several hundred neuromasts spread over the head, trunk, and tail fin. Lateral line neuromasts are mechanical low-pass filters that have an operating range from <1 Hz up to about 150 Hz. Within this frequency range, neuromasts encode the duration, local direction, amplitude, frequency, and phase of a hydrodynamic stimulus. This paper reviews the peripheral and central processing of lateral line information in fishes. Special attention is given to the coding of simple and complex hydrodynamic stimuli, to parallel processing, the roles of the various brain areas that process hydrodynamic information and the centrifugal (efferent) control of lateral line information. The review argues that in order to fully comprehend peripheral and central lateral line information processing, it is imperative to do comparative studies that take into account the ecology of fishes, meaning that natural stimulus and noise conditions have to be considered.     

Responses to dipole stimuli of anterior lateral line nerve fibres in goldfish, <Emphasis Type="Italic">Carassius auratus</Emphasis>, under still and running water conditions

We investigated how fibres in the anterior lateral line nerve of goldfish, Carassius auratus, respond to sinusoidal water motions in a background of still or running water. Two types of fibres were distinguished: type I fibres, which most likely innervate superficial neuromasts, were stimulated by running water (10 cm s⁻¹) while type II fibres, which most likely innervate canal neuromasts, were not stimulated by running water. The responses of type I fibres to sinusoidal water motions were masked in running water whereas responses of type II fibres were not masked. These findings are in agreement with previous data obtained from the posterior lateral line nerve of goldfish. Furthermore, we demonstrate here that for type I fibres the degree of response masking increased with increasing flow velocity. Finally, the ratio between responses that were masked in running water (type I) and those that were not masked (type II) increases with increasing flow velocity. Flow fluctuations that were generated by a cylinder in front of the fish did not affect ongoing activity in the flow, nor the dipole-evoked responses. The findings are discussed with respect to particle image velocimetry data of the water motions generated in the experiments.     

Development of free and canal neuromasts and their directions of maximum sensitivity in the larvae of ayu,Plecoglossus altivelis

Morphological changes in free neuromasts are reported from larvae of the Ayu,Plecoglossus altivelis. In newly-hatched larvae, free neuromasts were already recognizable in both the head and trunk. During larval growth, the number of free neuromasts increased, and the number of its sensory cells 2 days after hatching was constant. In the trunk, two types of free neuromasts, one with maximum sensitivity in the antero-posterior direction and the other with maximum sensitivity in the dorso-ventral direction, were observed. The former type predominated. In the head, free neuromasts were located around the eye and nose, their directions of maximum sensitivity forming lines tangential to concentric circles about the eye and nose. Distinct changes in free neuromasts occurred during the formation of the canal organ. The canal organ was first observed in the head region 64 days after hatching and in the trunk region 100 days after hatching. Concomitant with the formation of the canal organ, the profile of the cupulae of the free neuromasts changed from a flat bar to semispherical. Sensory cells in the canal neuromasts did not differ morphologically from those in the free neuromasts. It is considered that there is a close relationship between the sensitivity of the neuromast and the shape of the cupula, i.e., that the free neuromasts are adapted to slow water flow, as in lakes and the sea, while the neuromasts in the canal organ are adapted to rapid water flow.     

Water wave discrimination in the surface-feeding fish Aplocheilus lineatus

The surface-feeding fish Aplocheilus lineatus uses its cephalic lateral line to detect water surface waves caused by prey insects. The ability of Aplocheilus to discriminate between surface waves with aid of the lateral line system was tested by go/no-go conditioning. Our results show that Aplocheilus can distinguish between single-frequency surface wave stimuli with equal velocity or equal acceleration amplitudes which differ only in frequency. Frequency difference limens were about 15%, i.e. fish distinguished a 20-Hz wave stimulus from a 23-Hz stimulus in 100% of the trials. Aplocheilus can also discriminate between pure sine-wave stimuli and sine waves which show abrupt frequency changes. In contrast, fish were unable to distinguish amplitude-modulated wave stimuli (carrier frequency 20, 40 and 60 Hz, modulation frequency 10 and 20 Hz) from pure sine waves of the same frequency, even if amplitude modulation depth was 80%. Accepted: 27 December 1996