Comparative study on the morphology and the composition of the otoliths in the teleosts

Conclusion. Saccular otoliths of teleosts were mostly larger than utricular otoliths, which might relate to the three-dimensional movement. The large and heavy otolith may be better suited in saccules of the bottom and reef fishes. The quantities of iron in lagenar otoliths were found to be lower than those of birds. The function of the fish lagena remains to be elucidated by further studies. Objective. To evaluate the morphological characteristics and the chemical composition of the otoliths in fishes as related to behaviour and habitat. Materials and methods. We studied the morphology of the otoliths of 18 genera of fishes (81 samples) divided into 3 groups: saltwater fish (13 genera), freshwater fish except for the carp family (3 genera) and carp family fish (2 genera). The otoliths and the living environments were compared. The chemical composition was analysed using a synchrotron X-ray fluorescence analyser. Results. Bottom fishes generally have larger saccular otoliths, and migrating fishes have smaller saccular otoliths. In comparing the bottom/reef fishes and the migrating fishes in salt water, the former tended to have larger saccular otoliths. In saltwater bottom fishes the tendency was found that the thinner the head, the larger was the saccular otolith. We found significant quantities of iron, zinc and manganese in the lagenar otoliths.     

The relation between the migration function of birds and fishes and their lagenal function

CONCLUSION: The lagena of pigeons is a unique organ and it is concluded that it is a key element in the magnetic sensor system of pigeons and migrating birds. The lagenal otolith in pigeons contains more iron than saccular and utricular otoliths. The function of the lagena of pigeons was clarified because the homing ability of pigeons was largely disrupted after unilateral lagenal nerve section and attachment of magnetic balls with a magnetic field strength under 5 Gauss. The lagena of pigeons may have a navigational function as a geomagnetic sensor. OBJECTIVE: Otoliths of many kinds of fishes and birds were analyzed. MATERIALS AND METHODS: The otoliths of fish and birds were analyzed using synchrotron X-ray fluorescence analysis. Behavioral experiments concerning homing ability of pigeons were done by sectioning their lagenal nerves or interfering with the function of the lagena using a magnet. Twenty-one birds were treated in this way and 30 birds from the same loft of racing pigeons were used as controls. RESULTS: By comparing the compositions of the three different kinds of otoliths among several species of sea fish and birds, it was found that the saccular and utricular otoliths contain scarcely detectable levels of iron but that iron is present in significant quantities in the lagenal otoliths of the birds and sea fish. The results of homing tests clearly revealed a magnetic influence on the function of the lagena in terms of navigation ability of pigeons. The treated pigeons were either lost or significantly delayed while the controls returned within 30 min of release.     

Otolith regularities

The masses and the area sizes of the otoliths for the utriculus, sacculus and lagena of 15 species of the Black Sea fish are analyzed. Morphometrical otolith regularities are derived and their functional and ecomorphological explanations are suggested. The otolith regularities are summarized in four otolith rules: (1) the masses of the otoliths gradually increase with the fish growth. (2) The mass ratio of the sacculus and utriculus or the sacculus and lagena otoliths does not change with the fish growth. (3) The ratio between the otolith area s and the otolith mass m is described by the exponential equation s=alpham(2/3). (4) The ratio between the otolith and macula sizes does not change with fish growth. Mathematical modeling of the otolith displacement responses to the acoustic and the instant force stimuli is performed. Based on the modeling the functional and ecomorphological explanations of the otolith regularities are suggested: (1) the greater the otolith mass, the higher the acoustic sensitivity at low frequencies and the sharper the frequency-response curve at its maximum. (2) The separation between maxima of the frequency-response curves for the saccular and lagenar otoliths remains virtually constant with the fish growth. (3) The bottom and littoral fish have better auditory capabilities than the pelagic fish. (4) The sensitivity to vestibular stimuli for greater otoliths is higher but the response is slower. The corresponding acceleration resolution for greater otoliths is higher and the range of accelerations in which the otolith organ can operate is narrower. (5) The relative vestibular sensitivities of the utriculus, sacculus and lagena otolith organs remain constant with fish growth. (6) The otolith organs of the bottom and littoral fish are tuned to different accelerations and possess different functional properties. The otolith organs of pelagic fish are adapted to a limited range of accelerations and are less sensitive to low accelerations as compared to the bottom and littoral fish.     

Fish otolith mass asymmetry: morphometry and influence on acoustic functionality

The role of the fish otolith mass asymmetry in acoustic functionality is studied. The saccular, lagenar and utricular otoliths are weighted in two species of the Black Sea rays, 15 species of the Black Sea teleost fish and guppy fish. The dimensionless otolith mass asymmetry chi is calculated as ratio of the difference between masses of the right and left paired otoliths to average otolith mass. In the most fish studied the otolith mass asymmetry is within the range of -0.2 < chi < +0.2 (< 20%). We do not find specific fish species with extremely large or extremely small otolith asymmetry. The large otoliths do not belong solely to any particular side, left or right. The heavier otoliths of different otolithic organs can be located in different labyrinths. No relationship has been found between the magnitude of the otolith mass asymmetry and the length (mass, age) of the animal. The suggested fluctuation model of the otolith growth can interpret these results. The model supposes that the otolith growth rate varies slightly hither and thither during lifetime of the individual fish. Therefore, the sign of the relative otolith mass asymmetry can change several times in the process of the individual fish growth but within the range outlined above. Mathematical modeling shows that acoustic functionality (sensitivity, temporal processing, sound localization) of the fish can be disturbed by the otolith mass asymmetry. But this is valid only for the fish with largest otolith masses, characteristic of the bottom and littoral fish, and with highest otolith asymmetry. For most fish the values of otolith mass asymmetry is well below critical values. Thus, the most fish get around the troubles related to the otolith mass asymmetry. We suggest that a specific physicochemical mechanism of the paired otolith growth that maintains the otolith mass asymmetry at the lowest possible level should exist. However, the principle and details of this mechanism are still far from being understood.     

Magnetic Materials in Otoliths of Bird and Fish Lagena and Their Function

The mystery of the homing ability of pigeons has been the subject of much interest and it is widely believed that information from the earth's magnetic field may be involved. However, no specific magnetic sensory organ has yet been identified. The recent finding of magnetic materials in the lagenal otolith of fish and birds raises the possibility that these structures may be key elements in the elusive magnetic sensory system. For the elemental analysis of materials X-ray fluorescence using synchrotron radiation is one of the most powerful techniques available and was used in this study for analysis of the otoliths. By comparing the compositions of the three different kinds of otoliths among several species of sea fish and birds, we found that the saccular and utricular otoliths rarely contain detectable levels of iron but that iron is present in significant quantities in the lagenal otoliths of the birds. The lagenal otolith comprises tiny magnetic particles of low inertia that are displaced by imposed magnetic fields, providing the animal with geomagnetic sensory input, from which the brain would infer navigational information.     

Magnetic materials in otoliths of bird and fish lagena and their function

The mystery of the homing ability of pigeons has been the subject of much interest and it is widely believed that information from the earth's magnetic field may be involved. However, no specific magnetic sensory organ has yet been identified. The recent finding of magnetic materials in the lagenal otolith of fish and birds raises the possibility that these structures may be key elements in the elusive magnetic sensory system. For the elemental analysis of materials X-ray fluorescence using synchrotron radiation is one of the most powerful techniques available and was used in this study for analysis of the otoliths. By comparing the compositions of the three different kinds of otoliths among several species of sea fish and birds, we found that the saccular and utricular otoliths rarely contain detectable levels of iron but that iron is present in significant quantities in the lagenal otoliths of the birds. The lagenal otolith comprises tiny magnetic particles of low inertia that are displaced by imposed magnetic fields, providing the animal with geomagnetic sensory input, from which the brain would infer navigational information.     

Fish otolith asymmetry: morphometry and modeling

Mathematical modeling suggests that relatively large values of otolith mass asymmetry in fishes can alter acoustic functionality and may be responsible for abnormal fish behavior when subjected to weightlessness during parabolic or space flight [D.V. Lychakov, Y.T. Rebane, Otolith mass asymmetry in 18 species of fish and pigeon, J. Grav. Physiol. 11 (3) (2004) 17-34; D.V. Lychakov, Y.T. Rebane, Fish otolith mass asymmetry: morphometry and influence on acoustic functionality, Hear. Res. 201 (2005) 55-69]. The results of morphometric studies of otolith mass asymmetry suppose that the absolute value and the sign of the otolith mass asymmetry can change many times during the growth of individual fish within the range +/-20% [D.V. Lychakov, Y.T. Rebane, Otolith mass asymmetry in 18 species of fish and pigeon, J. Grav. Physiol. 11 (3) (2004) 17-34; D.V. Lychakov, Y.T. Rebane, Fish otolith mass asymmetry: morphometry and influence on acoustic functionality, Hear. Res. 201 (2005) 55-69]. This implies that the adverse effects of otolith asymmetry on acoustic and vestibular functionality could change during the lifetime of an individual fish. The aims of the present article were to examine the nature of otolith mass asymmetry fluctuation and to quantify otolith mass asymmetry in a large number of teleost fishes to verify our previous measurements. A dimensionless measure of otolith mass asymmetry, chi, was calculated as the difference between the masses of the right and left paired otoliths divided by average otolith mass. Saccular otolith mass asymmetry was studied in 59 Mediterranean teleost species (395 otolith pairs), 14 Black Sea teleost species (42 otolith pairs), red drum (196 otolith pairs) and guppy (30 otolith pairs). Utricular otolith mass asymmetry was studied in carp (103 otolith pairs) and goldfish (45 otolith pairs). In accordance with our previous results the value of chi did not depend on fish size (length or mass), systematic or ecological position of the fish, or otolith growth rate. In the great majority of the fishes studied, the saccular otolith chi was small /chi/ <0.05 (or <5%). Mathematical modeling indicates that values of chi vary among individual fish, but that the value is probably stable during a fish's lifetime.     

Appearance and evolution of calcitic and aragonitic otoconia during Pleurodeles waltl development.

The aim of this study is to determine the stages of appearance, morphology, crystallographic structure and chemical composition of otoconia during the inner ear development of an urodele amphibian, Pleurodeles waltl. The first otoconia are detected in the otocyst. Near hatching, calcitic otoconia are polyhedral in the saccule and cylindrical in the utricle. During the following stages, the saccular otoconia agglomerate and constitute a polyhedral calcitic otolith. At larval stage 44, aragonitic fusiform otoconia appear on the otolithic surface. At stage 52, X-ray diffraction analysis shows calcite and aragonite patterns. In adults, all the saccular otoconia are aragonitic. In contrast, the utricular otoconia do not show any modification up to adulthood. In the endolymphatic sac, otoconia appear at stage 45 and in the lagena at stage 49. They remain aragonitic up to adulthood. Energy dispersive X-ray spectroscopy (EDXS) elemental analysis of the otoconia reveals a high quantity of calcium with trace quantities of sodium, magnesium, phosphorus, sulfur, chlorine and potassium. However, magnesium and sulfur have a lower concentration in lagenar aragonitic otoconia than in utricular and saccular calcitic ones. As in adults, trace amounts of strontium are only found in aragonitic otoconia.     

Otolith mass asymmetries in the utricle and saccule of flatfish

The otolith mass of the saccules and utricles of plaice, Pleuronectes platessa (n = 39) and turbot, Psetta maxima (n = 21) was measured using an electronic microbalance. In the right-eyed plaice, the left utricular otoliths were found to be significantly heavier than the right (p < 0.0001), whereas no significant difference was found between left and right saccular otoliths (p < 0.751). In the left-eyed turbot, both the right utricular and saccular otoliths were found to be significantly heavier (in both cases, p < 0.0001). While the gene and regulative protein responsible for the peripheral biomineralisation process have been identified, it remains unclear how the symmetry between the right and left otoliths in fish species is regulated. Here it is likely that an additional central mechanism is involved. It must be assumed that similar processes govern the systematic asymmetry observed in flatfish such as the plaice and turbot. Taken together these findings are strongly suggestive of concomitant CNS modification and metamorphic plasticity, presumably represented in genetic code.     

On the origin of susceptibility to kinetotic swimming behaviour in fish: a parabolic aircraft flight study

Humans taking part in parabolic aircraft flights (PAFs) may suffer from motion sickness (SMS, a kinetosis; it comprises a dynamic and a static component). It has been argued that the so-called static variety of SMS during PAFs might be based on asymmetric statoliths (i.e., differently weighed statoliths on the right and the left side of the head), with asymmetric inputs to the brain being disclosed in microgravity. Since it has been repeatedly shown earlier that some fish of a given batch reveal a kinetotic behaviour during PAFs (especially so-called spinning movements and looping responses), we investigated whether fish swimming kinetotically in microgravity have a pronounced inner ear otolith asymmetry. Therefore, the swimming behaviour of larval cichlid fish was video-recorded during PAFs and subsequently, size and asymmetry (size difference between the left and the right side) of inner ear otoliths were determined. The asymmetry of utricular otoliths of kinetotic samples was found to be significantly higher than that of normally behaving experimental specimens. Regarding the asymmetry of saccular otoliths of the two groups, statistically different results were not obtained. The findings strongly support the earlier theoretical concept, according to which otolith asymmetry causes (static) SMS.     

Dendritic arbors on the saccule and lagena in the ear of the goldfish, Carassius auratus

The ear of the goldfish (Carassius auratus) contains three otolithic endorgans: the saccule, lagena, and utricle. The saccule has an auditory function in most teleost fishes for whom data are available, and there is evidence that the lagena is also an auditory endorgan in the goldfish. This study was conducted to compare the innervation of the saccule and the lagena to one another and to previously published data from goldfish and other species. We placed cobaltous-lysine in saccular and lagenar nerves in vivo and permitted uptake over 18-24 h. A total of 59 saccular and 59 lagenar dendritic arbors were labeled in 10 fishes. Our data indicate that arbors on the saccule and lagena have similar morphologies, but differ in relative size. Saccular arbors tend to be smaller than lagenar arbors, with median arbor widths of 50 micrometer on the saccule and 74 micrometer on the lagena. Fiber diameters on the two endorgans are similar. A regional analysis of the saccule indicated that a wide range of arbor sizes are found along the rostral-caudal axis, with larger arbors more common caudally. Our data do not support the presence of two distinct categories of saccular afferents with non-overlapping distributions. Moderate arbor widths (50-99 micrometer) were most common in all regions of the lagena. Maximum arbor width and hair cell density do not appear to be correlated with one another on either the saccule or the lagena. Comparisons with published data from goldfish and oscar revealed similarities and differences that may be attributable to variations in label uptake or transport as well as potential species differences.     

前庭器官超微结构研究及结构图构建

目的通过扫描电镜观察小鼠椭圆囊、球囊、壶腹超微结构,研究分析并据此构建前庭器官新的结构示意图。方法选取3个年龄段小白鼠各10只,分别是年轻组≤2个月、中年组2~12个月、老龄组>12个月。分离出椭圆囊、球囊、壶腹,采用扫描电镜技术样品制备方法制备样品,应用扫描电镜进行样品观察。结果扫描电镜下可以得到:①椭圆囊斑及球囊斑不同层面图片:表面为堆积并相互黏附的"表面耳石",表面耳石下是无结构胶状质;底层表面耳石深入到无结构胶质层里;无结构胶质层下面是毛细胞纤毛及"纤毛间耳石"层,不同纤毛束之间均有纤毛间耳石存在,立于支持细胞表面,表面平坦;蜂窝状胶质物质联接无结构胶质层、纤毛间耳石及毛细胞纤毛。②壶腹超微结构的图片:嵴帽是无结构的胶状质与壶腹外侧壁紧贴,但较易分离,嵴帽和壶腹外侧壁之间有纤细的晶状体物质,在壶腹嵴两侧壁上也有纤细晶体物质(壶腹嵴表面耳石);不同的毛细胞纤毛之间有耳石结构的存在(壶腹嵴纤毛间耳石)。结论通过对前庭器官扫描电镜的观察,发现了椭圆、球囊斑及壶腹的新结构成分,由此构建出新的前庭器官超微结构示意图。     

Organization and connections of the dorsal descending nucleus and other presumed acoustic areas in the brainstem of the teleost fish, Astronotus ocellatus

This study provides new information on brainstem areas, assumed to be auditory based on observations in other species, in the oscar, Astronotus ocellatus. The primary goal of the study was to explore the morphology of the dorsal descending octaval nucleus, which contains a population of neurons that receives acoustic afferents from the inner ear. Using cytoarchitectonic and connectional criteria, we revised the previously defined dorsal boundary of the descending octaval nucleus, such that the most dorsomedial neurons in this nucleus are positioned ventral to the cerebellar crest and medial to nucleus medialis. At some levels, these dorsomedial cells are continuous with another part of the dorsal descending nucleus that underlies nucleus medialis. The terminal fields of the saccule and lagena are located within this latter, more ventral part of the dorsal descending nucleus. However, the dorsomedial cells that are proximate to the cerebellar crest have long ventral dendrites that extend into these terminal fields, and therefore likely receive saccular and lagenar input. In contrast to a previous report, saccular afferents terminate more medially within the dorsal descending nucleus than do lagenar inputs. Injections of horseradish peroxidase in nucleus centralis of the torus semicircularis revealed that many descending nucleus neurons that lie within the saccular and lagenar terminal fields, including the dorsomedial neurons proximate to the cerebellar crest, project to this acoustic midbrain area. These injections also revealed a secondary octaval population like that described in otophysan fishes.     

A critical period for gravitational effects on otolith formation

The otoliths of adult animals do not change significantly during space flight. However, during the period when otoliths are first developing, rearing in space produces significantly larger otoliths. Conversely, animals reared on a centrifuge have smaller than normal otoliths. To identify a critical period during development for gravitational effects on otolith growth, fertilized zebrafish (Danio rerio) eggs were reared on a centrifuge for 1 week. The fine structure of their inner ear during development was studied by both light- and transmission electron microscopy. By 16 hours after fertilization (1-g, at 28.5 degrees C), precursors of the otoliths are seen but no sign of a sensory epithelium is present. Mature hair cells, appearing capable of mechanotransduction, are not seen until between 48 and 72 hours after fertilization. Zebrafish reared at 3-g from 1 to 7 days after fertilization exhibit significantly slower otolith growth than did 1-g controls. Fish exposed to 3-g only from 12-36 h after fertilization had slightly smaller otoliths than 1-g controls, but this difference was not significant. Animals exposed to 3-g from 36 h to 7d after fertilization did have significantly smaller otoliths. If the fish use their hair cells to assess otolith weight in a regulatory role, the hair cells would have to be functional. Thus the earliest stage zebrafish, which were not significantly affected by centrifugation, probably did not have an adequate means of sensing otolith weight to "correct" for the excess weight.     

Input–Output Functions of Vestibular Afferent Responses to Air-Conducted Clicks in Rats

Sound-evoked vestibular myogenic potentials recorded from the sternocleidomastoid muscles (the cervical vestibular-evoked myogenic potential or cVEMP) and the extraocular muscles (the ocular VEMP or oVEMP) have proven useful in clinical assessment of vestibular function. VEMPs are commonly interpreted as a test of saccular function, based on neurophysiological evidence showing activation of saccular afferents by intense acoustic click stimuli. However, recent neurophysiological studies suggest that the clicks used in clinical VEMP tests activate vestibular end organs other than the saccule. To provide the neural basis for interpreting clinical VEMP testing results, the present study examined the extent to which air-conducted clicks differentially activate the various vestibular end organs at several intensities and durations in Sprague–Dawley rats. Single unit recordings were made from 562 vestibular afferents that innervated the otoliths [inferior branch otolith (IO) and superior branch otolith (SO)], the anterior canal (AC), the horizontal canal (HC), and the posterior canal (PC). Clicks higher than 60 dB SL (re-auditory brainstem response threshold) activated both semicircular canal and otolith organ afferents. Clicks at or below 60 dB SL, however, activated only otolith organ afferents. Longer duration clicks evoked larger responses in AC, HC, and SO afferents, but not in IO afferents. Intra-axonal recording and labeling confirmed that sound sensitive vestibular afferents innervated the horizontal and anterior canal cristae as well as the saccular and utricular maculae. Interestingly, all sound sensitive afferents are calyx-bearing fibers. These results demonstrate stimulus-dependent acoustic activation of both semicircular canals and otolith organs, and suggest that sound activation of vestibular end organs other than the saccule should not be ruled out when designing and interpreting clinical VEMP tests.     

Growth of a fish ear: 1. Quantitative analysis of hair cell and ganglion cell proliferation

Proliferation (or addition) of inner ear sensory hair cells continues for a long time postembryonically in cartilaginous and bony fishes, and in amphibians. In contrast, proliferation only occurs during embryonic development in birds and mammals. However, detailed quantitative data on hair cell addition are not available for bony fishes. In order to quantify the extent of proliferation, we determined the number of sensory hair cells on the saccular sensory epithelium in specimens of the cichlid fish Astronotus ocellatus (the oscar) ranging from 2.0 to 19.0 cm in standard length (0.9-343 g). Ganglion cells were counted using serial sections of the saccular branch of the eighth nerve in animals of the same size range. The saccular macula of a 2.0 cm long (0.9 g) Astronotus contains approximately 5500 sensory hair cells; fish from 16 to 19 cm long have over 170 000 hair cells. The increase in number of sensory cells and the increase in both length and weight of the animals studied were statistically correlated (r2 = 0.8). The relative densities of saccular sensory cells in different epithelial regions remained constant in animals from 2.0 to 17 cm; in larger animals the cell density decreased somewhat. Based upon very conservative estimates of the rate of growth of Astronotus, we calculate that an average of 167 hair cells/day are added during the time when the cell population of the saccule increases. Ganglion cell number also increased approximately 4.8 times in the range of fish studied. The smallest animals in our study had about 150 ganglion cells per saccular epithelium, while the largest fish had over 600 ganglion cells. We estimate that the average ratio of hair cells to afferent fibers increases from about 30:1 in the smallest fish to over 300:1 in the largest animals.     

Models of the mechanical sensitivity and growth of otoliths in fish

It has been suggested that, in the fish, the change of otolith mass during development under altered gravity conditions and the growth of otoliths in normal conditions, are determined by feedback between otolith dynamics and the processes that regulate otolith growth. The hypothesis originates from an oscillator model of the otolith in which otolith mass is one of the parameters. However, the validity of this hypothesis is not obvious and has not been experimentally verified. We tested this hypothesis by comparing the oscillator model with a simplified spatially distributed model of the otolith. It was shown that in the case of a spatially distributed fixation of the otolith plate (otoconial layer) to the macular surface, the mechanical sensitivity of the otolith does not depend on the total otolith mass nor on its longitudinal size. It is determined by otolith thickness, the Young's modulus and viscosity of gel layer of the growing otolith. These parameters may change in order to maintain otolith sensitivity under conditions (such as growth or altered gravity) that change the dynamics of otolith movement.     

Sparc Protein Is Required for Normal Growth of Zebrafish Otoliths

Otoliths and the homologous otoconia in the inner ear are essential for balance. Their morphogenesis is less understood than that of other biominerals, such as bone, and only a small number of their constituent proteins have been characterized. As a novel approach to identify unknown otolith proteins, we employed shotgun proteomics to analyze crude extracts from trout and catfish otoliths. We found three proteins that had not been associated previously with otolith or otoconia formation: ‘Secreted acidic cysteine rich glycoprotein’ (Sparc), an important bone protein that binds collagen and Ca²⁺; precerebellin-like protein, which contains a C1q domain and may associate with the collagenous otolin-1 during its assembly into a framework; and neuroserpin, a serine protease inhibitor that may regulate local protease activity during framework assembly. We then used the zebrafish to investigate whether Sparc plays a role in otolith morphogenesis. Immunodetection demonstrated that Sparc is a true constituent of otoliths. Knockdown of Sparc expression in morphant zebrafish resulted in four principal types of defective otoliths: smaller, extra and ectopic, missing and fused, or completely absent. Smaller size was the predominant phenotype and independent of the severity of otic-vesicle defects. These results suggested that Sparc is directly required for normal otolith growth.     

Modeling the relation between head orientations and otolith responses in humans

We have performed a finite element simulation of realistic displacements of otolith membranes by static linear accelerations. The simulations were based on accurate measurements of the surfaces of human utricular and saccular maculae, which indicate a clear curvature of these surfaces. The results show that this curvature, a feature probably found in all mammals, has no effect on the mechanics of the structure as a whole since the elastic coupling in the otolith membrane is insufficient. Hair cell excitations on any place of the macula are only affected by the local orientation of the macula with respect to acceleration. Based on the displacements of the otolith membrane, we also calculated the induced activation patterns on the otolith epithelia. These patterns provide for the first time a complete image of peripheral otolith activity. The individual activation patterns at selected locations on the macula correspond well with single cell recordings of actual peripheral otolith neurons.     

Morphometry of otoconia in the utricle and saccule of developing Japanese quail

The development of otoconia in the utricular and saccular maculae from initial embryonic formation to adult stages was examined in Japanese quails. Both the morphology and size of the otoconia were quantified at different developmental stages. It was observed that the otoconia were initially formed on embryologic stage E5 in the saccule and E6 in the utricle. Otolith mass areas increased in a sigmoidal growth pattern, with saccular otolith areas being smaller than the utricular mass areas. Saccular otolith masses reached adult values at embryonic stage E12 and utricular areas reached adult values at post-hatch day 7. Mature individual otoconia were characterized by a barrel shape with two trihedral faceted ends. However, initial formation of otoconia at E5 (saccular) and E6 (utricular) maculae was characterized by a double fluted morphology that consisted of an hourglass shape with extended fins forming trihedral angles of 120 degrees. Double fluted otoconia rapidly filled, so that by embryonic day 8 mature otoconia dominated the maculae for the remainder of development through adulthood. Thus, a progression from double fluted to mature forms was noted. Mature utricular otoconia in adult quails averaged 11 microm in length and 5 microm in width, with length/width ratios of approximately 2.5:1, for all size ranges. Saccular otoconia were smaller, having about 70% the size of utricular otoconia in both length and width. During development, the average size and range of individual otoconia increased nearly linearly for both otolith organs. In the utricular macula, large otoconia were concentrated in the lateral regions of the epithelium. In contrast, otoconia of various sizes were distributed uniformly across the surface of the saccular macula.