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.     

The responses of primary and secondary sensory hair cells in the squid statocyst to mechanical stimulation

Summary Intracellular recordings were obtained from primary and secondary sensory hair cells in the anterior transverse crista segment of the squid (Alloteuthis subulata) statocyst during imposed displacements of the overlying cupula. The secondary sensory hair cells were depolarized by ventral movements of the cupula and hyperpolarized by dorsal cupula movements. The displacement/response curve was asymmetric around the zero position and sigmoidal in shape, similar to that already described for vertebrate hair cells. The cells are estimated to have a sensitivity of at least 0.5 mV per degree angle of cilia displacement. The responses showed pronounced adaptation and could be blocked by bath applied alcohols, such as heptanol or octanol, or by high concentrations of aminoglycosides.The primary sensory hair cells were depolarized by dorsal movements of the cupula, usually responding with a burst of action potentials. The displacement/response curve was also sigmoidal in shape and the firing pattern showed strong adaptation to maintained displacements of the cupula.The cupula itself appeared to be irregular in shape, extending much further into the statocyst cavity in its central part than at its edges. This is likely to result in differences in the responses of the underlying hair cells along the length of the crista ridge.     

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.     

Experimental measurement of the stiffness of the cupula.

An experimental procedure is described which consists of cutting the canal duct, inserting a micropipette and administering known volumetric displacements to the cupula. The cupula is made visible by dying the endolymph. Known displacements are administered to the cupula, and the time constant of the return to its equilibrium position is measured. With this information, the stiffness of the cupula is calculated. The experiment was successfully carried out on five White King pigeons. The mean stiffness found in somewhat less than other results reported in the literature, and reasons for this discrepancy are noted.     

Side specific effect of 5-hydroxytryptamine on NaCl transport in the apical and basolateral membrane of rat tracheal epithelia

5-Hydroxytryptamine (5-HT) can be released from mast cells and platelets through an IgE-dependent mechanism and may play a role in the pathogenesis of allergic bronchoconstriction. However, the effect of 5-HT on ion transport by the airway epithelium is still controversial. The objective of this study was to determine whether 5-hydroxytryptamine (5-HT) regulates NaCl transport by different mechanisms in the apical and basolateral membrane of tracheal epithelia. We studied the rat tracheal epithelium under short-circuit conditions in vitro. Short-circuit current (I(sc)) was measured in rat tracheal epithelial monolayers cultured on porous filters. 5-HT inhibited Na(+) absorption [measured via Na(+) short-circuit current (I(Na)(sc))] in the apical membrane and stimulated Cl(-) secretion [measured via Cl(-) short-circuit current (I(Cl)(sc))] in the basolateral membrane. Functional localization using selective 5-HT agonists and antagonists suggest that I(Cl)(sc)is stimulated by the basolateral membrane-resident 5-HT receptors, whereas I(Na)(sc) is inhibited by the apical membrane-resident 5-HT2 receptors. The basolateral addition of 5-HT increases intracellular cAMP content, but its apical addition does not. The addition of BAPTA/AM blocked the decrease of I(Na)(sc)which was induced by the apical addition of 5-HT, and 5-HT increased intracellular Ca concentrations. These results indicate that 5-HT differentially affects I(Na)(sc)and I(Cl)(sc)across rat tracheal monolayers through interactions with distinct receptors in the apical and the basolateral membrane. These effects may result in an increase of water movement towards the airway lumen.     

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.     

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.     

A mouse model for benign paroxysmal positional vertigo with genetic predisposition for displaced otoconia

Abnormal formation of otoconia, the biominerals of the inner ear, results in balance disorders. The inertial mass of otoconia activates the underlying mechanosensory hair cells in response to change in head position primarily during linear and rotational acceleration. Otoconia associate exclusively with the two gravity receptors, the utricle and saccule. The cristae sensory epithelium is associated with an extracellular gelatinous matrix known as cupula, equivalent to otoconia. During head rotation, the inertia of endolymphatic fluids within the semicircular canals deflects the cupula of the corresponding crista and activates the underlying mechanosensory hair cells. It is believed that detached free‐floating otoconia particles travel ectopically to the semicircular canal and cristae and are the culprit for benign paroxysmal positional vertigo (BPPV). The Slc26a4 mouse mutant harbors a missense mutation in pendrin. This mutation leads to impaired transport activity of pendrin and to defects in otoconia composition and distribution. All Slc26a4 ^loop/loop homozygous mutant mice are profoundly deaf but show inconsistent vestibular deficiency. A panel of behavioral tests was utilized in order to generate a scoring method for vestibular function. A pathological finding of displaced otoconia was identified consistently in the inner ears of mutant mice with severe vestibular dysfunction. In this work, we present a mouse model with a genetic predisposition for ectopic otoconia with a clinical correlation to BPPV. This unique mouse model can serve as a platform for further investigation of BPPV pathophysiology, and for developing novel treatment approaches in a live animal model.     

Effect of hypothyroidism on responsiveness to beta-adrenergic stimulation.

Chronic administration of aminotriazole (0.5 g/kg food) to rats was accompanied by a reduced responsiveness to acute administration of the beta-adrenergic agonist, l-isoproterenol (50-100 mug/kg, sc). The responses tested included water intake, change in heart rate in the anesthetized and unanesthetized rat, change in mean blood pressure, and change in blood glucose concentration. In addition, the increase in tail skin temperature accompanying administration of epinephrine (1 mg/kg, sc) was significantly reduced in the hypothyroid group. Administration of l-thyroxine (25 mug/kg per day, ip) to aminotriazole-treated rats prevented the reduction in responsiveness to beta-adrenergic stimulation. Thus, an interaction appears to exist between the level of thyroid activity and responsiveness to beta-adrenergic agonists in rats.     

Functional morphology of the crista ampullaris: with special interests in sensory hairs and cupula: a review.

The functional significance of the ciliary interconnections and cupula has been reviewed. The ciliary interconnecting systems are divided into 2 types, i.e. side links and tip links. The side links acts to maintain the regular distance between the cilia thereby keeping the geometrical arrangement of the entire sensory hair bundle intact as well as to prevent close contact between neighbouring cilia. The tip links, stretching upwards from the tips of the shorter stereocilia to their taller neighbouring shafts, are actually involved in mechanoelectrical transduction. The cupula is composed of the cupula and subcupular meshwork. The subcupular meshwork consists of long branching filaments cross-bridged to one another. The cupula would function as a rigid plate and equally distribute the shear force of the cupula to all the ciliary bundles. The subcupular meshwork may play a role in the transmission of the shear strain force of the cupula to the ciliary bundle and may also exert an additional damping effect in order to prevent unwanted vibrations.     

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.     

Some aspects of the gross and fine structure of the amphibian papilla in the labyrinth of the newt, Triturus cristatus

Each receptor cell in the sensory macula bears a number of stereocilia and one peripherally located kinocilium; in the two halves of the macula, the kinocilia lie on opposite sides of their associated stereocilia. The morphological axes of the receptor cells are approximately parallel to the long axis of the papilla. The gelatinous cupula overlying the macula extends almost to the opposite wall of the papilla. These structural features are discussed in connection with both the proposed function of the papilla as a vibration detector and the possible evolutionary relationships with other acousticolateralis receptors.     

Structures transmitting stimulatory force to the sensory hairs of vestibular ampullae of fishes and frog

Summary Serial sections of the vestibular ampullae of two species of fish and one species of frog were investigated by electron microscopy. The kinocilium is the only connection between the sensory cells and the auxiliary structure (cupula). The cupula possesses canals that traverse its entire height. Each canal contains a single kinocilium in its proximal part; distally, it is filled with material that stains with colloidal silver. The matrix of the cupula consists of filaments running perpendicular to the canals. These filaments do not stain with colloidal silver. The kinocilium is connected to the wall of the canal via structures that differ in the studied species of fish and frog. The filamentous links between the kinocilium and the longest stereovilli of the sensory hair bundle are similar in all the investigated species. The stereovilli are interconnected by basal and shaft links, and by horizontal and oblique tip connectors, similar to those described by other authors for macula organs and the organ of Corti, although differences in structural details, especially of the horizontal tip and the shaft connectors, are present. Some of these are species specific and some are related to the position of the sensory cell in the epithelium and/or specific to the organ (ampulla or macula organ). Some attachment sites of the links are associated with osmiophilic submembranous material. These differences in the structure, distribution and attachment sites of the links are possibly of functional importance.     

Effect of temperature on auditory receptor functions in bushcrickets (Orthoptera, Tettigonioidea)

By electrophysiological methods, effect of temperature on bushcricket tympanal organ functions was studied. Activity of auditory receptors was recorded intracellularly in the 5th nerve of I thoracic ganglion in Tettigonia cantans, Metrioptera roeselii, M. bicolor, Platycleis albopunctata, Pholidoptera griseoaptera, and Phaneroptera falcata. The temperature was changed in the range from 17 to 34 degrees C. Heating of the tympanal organ to 30-32 degrees C led to a decrease of impulse amplitude, shortening of their duration, an increase of sensitivity, of the burst instantaneous frequency, and of the number of impulses in responses as well as to a decrease of latent periods (LP) of receptor reaction. The optimal frequency in all studied cells did not change, although range of perceived frequencies was enlarged. The frequency threshold curve of receptors either was shifted down along the ordinate scale without changes of its shape or the thresholds at various frequencies decreased non-uniformly. Thus, the obtained data indicate the absence of changes in the frequency tuning of the auditory receptors with changes of temperature.     

Hibernation and daily torpor in an armadillo, the pichi (Zaedyus pichiy).

Hibernation and daily torpor are physiological strategies to cope with energetic challenges that occur in many mammalian and avian taxa, but no reliable information exists about daily torpor or hibernation for any xenarthran. Our objective was to determine whether the pichi (Zaedyus pichiy), a small armadillo (Xenarthra, Dasypodidae) that inhabits arid and semi-arid habitats in central and southern Argentina and Chile, enters shallow daily torpor or prolonged deep hibernation during winter when environmental temperature and food availability are low. We studied body temperature changes during winter in semi-captive pichis by means of temperature dataloggers implanted subcutaneously. All individuals entered hibernation, characterized by torpor events of 75+/-20 h during which the subcutaneous temperature (T(sc)) decreased to 14.6+/-2.1 degrees C. These events were interrupted by periods of euthermia of 44+/-38 h with a T(sc) of 29.1+/-0.7 degrees C. After the hibernation season, daily torpor bouts of 4 to 6 h occurred irregularly, with T(sc) dropping to as low as 24.5 degrees C. We conclude that the pichi is a true hibernator and can enter daily torpor outside of the hibernation season.     

Differences in action of topical and systemic cysteamine on gastric blood flow,gastric acid secretion and gastric ulceration in the rat

The effect of cysteamine on gastric blood flow and on the indomethacin-induced gastric mucosal damage was studied. In anesthetized rats, cysteamine (280 mg/kg) given subcutaneously (sc) decreased gastric blood flow measured by the laser Doppler flowmetry technique. In contrast, cysteamine (1–60 mg/ml) applied topically to the serosal surface of the stomach evoked a concentration-dependent and long-lasting increase in gastric blood flow. At 60 mg/ml, cysteamine increased blood flow by 166.8 ± 26.1% of predrug control value. Pretreatment with indomethacin (20 mg/kg, sc), intravenous (iv) atropine (1 mg/kg), propranolol (1 mg/kg, iv), combined H₁ and H₂-blockade or bilateral cervical vagotomy alone or combined with iv guanethidine (8 mg/kg), or pretreatment with the capsaicin analogue resiniferatoxin did not reduce the vasodilator response to cysteamine. The vasodilator response to topical capsaicin, was not reduced after sc cysteamine (280 mg/kg) pretreatment. In conscious pylorus-ligated rats, sc cysteamine (100 or 280 mg/kg) given simultaneously with indomethacin inhibited gastric acid output but had variable effects on the indomethacin-induced gastric mucosal damage. Cysteamine (100 or 280 mg/kg) administered sc 4 h prior to indomethacin enhanced gastric injury by sc indomethacin, but did not prevent the gastroprotective action of capsaicin. In contrast, orally administered cysteamine (60 mg/ml) reduced gastric injury induced by sc indomethacin plus intragastric HCl. These data provide the first evidence for the effect of cysteamine on gastric microcirculation in the rat and suggest a direct vasodilator effect for topical cysteamine. The microvascular effects of cysteamine are largely responsible for the different effects of this agent on experimental gastric injury.     

Temporal relationships between warmth imagery and associated changes in digital pulse amplitude, skin temperature and skin temperature sensation

Temporal relationships between warmth imagery and associated psychophysiological changes were studied by recording digital pulse amplitude, skin temperature, and thermal sensations in 5 subjects. After each trial the subjects were asked whether they thought they had been successful in producing the expected mental image. During the subjectively successful imagery tasks the digital pulse amplitude and the skin temperature of the hand rose significantly. The subjective onset of imagery took place after the digital pulse amplitude had started to change but before the skin temperature had begun to rise. This implies that mental imagery of skin warming as a conscious experience is not a prerequisite of somatic change. The thermal sensation (i.e. feeling of warmth) took place while the skin temperature was rising or immediately after that. This suggests that it is not caused by the mental image per se but by activation of skin temperature receptors.     

Effect of temperature and monensin on the level of intracellular sodium and pH in perfused RIF-1 cultured cells

The direct measurement of temperature in subcutaneously (sc) implanted tumors shows that the actual tumor temperature is by 3-4 degrees C lower than the normal body temperature. Thus, the temperatures usually used for tumor hyperthermia are in fact heating the sc-tumors from 33 to 37 degrees C. The temperature increase during the perfusion of radiation-induced fibrosarcoma (RIF-1) cells from 33 to 37 degrees C caused a reversible increase in intracellular 23Na ([Na+]i) NMR signal intensity by 50-60%. This heating significantly decreased the intracellular pH (pH) in 5 min, but it returned back to the baseline level during the heating period. The 3ATP/P(i) remained generally unchanged throughout the experiment. Monensin, an antitumoral drug and Na+ ionophor, increased [Na+]i by 20% during cell superfusion without heating. When combined, monensin did not increase the heating effect on [Na+]i. However, when monensin was added to the superfusion media, the [Na+]i level did not return to baseline during post-heating recovery, but instead started to increase again. Monensin did not significantly change pH(i) and betaATP/P(i). Our data and the literature show that monensin can accelerate the processes leading to the collapse of the transmembrane Na+ gradient and thus can increase the thermo-sensitivity of tumor cells.     

外加直流电场对血管平滑肌细胞形态及迁移行为的影响

在外加直流电场(electricalfields,EFs)作用下血管平滑肌细胞(VSMCs)膜表面细胞生长因子受体表达发生明显的变化,并影响细胞形态、迁移的特性。通过EFs干预装置干预体外培养的大鼠主动脉VSMCs,记录和分析细胞图像,研究不同强度电场、不同作用时间下VSMCs迁移和细胞形态的变化,并用免疫细胞化学或免疫荧光染色方法检测与VSMCs迁移相关的血小板衍化生长因子受体(PDGFR)、血管紧张素II1型受体(AT1R)和2型受体(AT2R)等受体的表达情况,研究EFs影响VSMCs形态及迁移的机制。研究结果提示,在EFs干预作用下,VSMCs膜PDGFR表达增加,部分细胞呈不对称分布,在EFs阴极面较集中;细胞中AT1R表达亦增加,但无明显不对称分布现象;AT2R表达没有改变;EFs长时间作用下,培养的VSMCs有明显的电场趋化性,细胞向阴极迁移的距离明显高于无EFs作用对照组,细胞膜向阴极方向伸展,发生形状改变,定向迁移依赖于EFs强度。EFs作用下,部分细胞生长因子受体的表达上调和重分布,可能与细胞定向迁移的启动和维持有关。     

Effect of l-menthol on laryngeal receptors

We have studied the effect of l-menthol on laryngeal receptors. Experiments have been conducted in 11 anesthetized dogs that breathed through a tracheostomy. We have recorded the activity of 23 laryngeal cold receptors and 19 mechanoreceptors. Constant flows of air, 15-50 ml/s (low) and 100-150 ml/s (high), passing for 10 s through the isolated upper airway in the expiratory direction, lowered laryngeal temperature and activated the cold receptors. This cold-induced discharge promptly ceased upon withdrawal of the airflow. Addition of l-menthol to the airflow evoked, for a similar decrease in temperature, a greater peak activation of the cold receptors than airflow alone (low flows 164%, high flows 111%); statistical significance was reached only for the lower flow. This activity outlasted the cessation of airflow by 30-120 s, even at a time when laryngeal temperature had returned to control (low flow 237%, high flow 307% of similar trials with airflow alone). Four laryngeal cold receptors were also tested with l-menthol added to a warm, humidified airflow that did not change laryngeal temperature; all of them were stimulated with a long-lasting discharge. Nine cold receptors were also tested with d-neomenthol and d-isomenthol; both isomers stimulated the receptors. None of the 19 mechano-receptors tested was affected by l-menthol. We conclude that l-menthol constitutes a specific stimulant of laryngeal cold receptors and could provide a useful tool for the study of their reflex effects.