Morphological and fine structural features of the basilar papilla in ambystomatid salamanders (Amphibia; Caudata)

The morphology and fine structure of the basilar recess and basilar papilla were investigated in four species of salamanders from the family Ambystomatidae. The otic relationships of the recess and papilla to the proximal part of the lagena and saccule are described, and new terminology is suggested for the periotic relationships of the basilar recess to a diverticulum of an intracapsular periotic sac. The basilar papilla consists of supporting cells united laterally by gap junctions, capped by microvilli uniformly arranged around a short, central cilium, and hair cells that typically show several synapses with a single afferent nerve fiber, each marked by a rounded synaptic body surrounded by vesicles. In contrast to anuran basilar papillae, efferent nerve terminals were observed in synapse with hair cells and, rarely, upon afferent fibers. The distal half of the ambystomatid papilla contained hair cells capped by tall ciliary bundles, with kinocilia that show swellings near their tips with delicate attachments to adjacent tall stereocilia. A tectorial body covers only this region of the papilla. Hair cells with shorter stereocilia, situated in the proximal half and at the papillar margins, are related only to filamentous extensions of the tectorial body. The ambystomatid basilar recess and papilla are compared to auditory end-organs in other vertebrates, and it is suggested that a basic distinction can be made between aural neuroepithelia in amniotes versus that in nonamniotic vertebrate ears.     

Ultrastructure of serotonin-containing cells in the pineal organ of Lampetra planeri (Petromyzontidae)

Summary The ultrastructure of the cells containing residual bodies (Collin, 1969) was investigated in the pineal organ of Lampetra planeri. These cells are characterized by their indoleamine metabolism (Meiniel, 1978; Meiniel and Hartwig, 1980). Morphologically, they belong mainly to two types: (1) a photoreceptor cell type, and (2) a pinealocyte cell type. The first type is present in the pineal sensory epithelium and in the atrium, while the second is observed in the deep part of the atrium. Intermediate cell types are rare. All these cells are characterized by the presence of voluminous dense bodies, the 5-HT-storing structures, in their cytoplasm.The elongated cone-type photoreceptor cells show a segmental organization and well-developed outer segments consisting of short disks (2–3 m), while their basal pedicles form synapses with the dendritic processes of neurons. The pinealocytes are spherical or oval in shape, their receptor poles being regressed to cilia of the 9+0 type. In these cells, no synaptic ribbons have to date been observed. In both cell types a Golgi apparatus is present producing dense granules 130 nm in diameter and a polymorphous dense material.The photoreceptor cells most probably respond to light and transmit a sensory (i.e., nervous) message. In addition, they produce and metabolize indoleamines, probably including, melatonin (Meiniel, 1978; Meiniel and Hartwig, 1980). The pinealocytes, in spite of their loss of direct photosensitivity, retain their capacity to metabolize indoleamines (Meiniel, 1978; Meiniel and Hartwig, 1980).The presence, in the same pineal organ, of another photoreceptor cell type (cf. Collin, 1969–1971) differing morphologically as well as biochemically (no detectable indoleamine metabolism) from the photoreceptor cell type described in the present investigation, points to the existence of two different sensory cell lines: (1) a pure photoreceptor line, and (2) a photoneuroendocrine line. The phylogenetic evolution of these two cell lines is discussed in terms of functional analogy.     

Scanning electron microscope studies of the papilla basilaris of some turtles and snakes

The papillae basilares of three species of turtles and four species of snakes were studied by SEM. The papillae of turtle are relatively large among reptiles and are characterized by a long, horizontal middle section resting on wide basilar membrane. Both terminal ends of the papilla extend onto the surrounding limbus in the form of a forked or "T" -shaped end or as a curved, "hook"- like processes. Details vary with the species. In the three species of turtles studied, there were between 1,100 and 1,400 hair cells on a papilla. The tectorial membrane covering the horizontal portion of the papilla is heavy in appearance and tightly attached to the kinocilial bulbs. The terminal ends of the papilla are covered by a thin gelatinous material. In addition, mat-like tectorial network covers the supporting cells and extends from the microvilli of the supporting cells to the overlying tectorial membrane. All hair cells are unidirectionally and abneurally oriented. The supporting cell surfaces form a large part of the papilla and, thus, hair cell density is low. The papillae of the two boid snake species studied are moderately long among snakes and contain a moderate number of hair cells (574 in Epicrates and 710-780 in Constrictor). Papillar form is elongate, avoid, or canoe-shaped. The tectorial membrane may be either highly fenestrated or moderately dense and covers all but a few of the terminal hair cells. A tectorial-like mat covers all but a few of the terminal hair cells. Most hair cells are unidirectionally and abneurally oriented. A few terminal cells in boids may show reverse orientation. Hair cell density is similar to that of turtles.     

The ultrastructure and innervation of the ear of the gar, Lepisosteus osseus

The endorgans of the inner ear of the gar were examined using transmission and scanning electron microscopy as well as nerve staining. The ultrastructure of the sensory hair cells and supporting cells of the gar ear are similar to cells in other bony fishes, whereas there are significant differences between the gar and other bony fishes in the orientations patterns of the sensory hair cells on the saccular and lagenar sensory epithelia. The saccular sensory epithelium has two regions, a main region and a secondary region ventral to the main region. The ciliary bundles on the main region are divided into two groups, one oriented dorsally and the other ventrally. Furthermore, as a result of curvature of the saccular sensory epithelium, the dorsal and ventral ciliary bundles on the rostral portion of the epithelium are rotated ninety degrees and are thus oriented on the animal's rostro-caudal axis. Hair cells on the secondary region are generally oriented ventrally. The lagenar epithelium has three groups of sensory hair cells. The groups on the rostral and caudal ends of the macula are oriented dorsally, whereas the middle group is oriented ventrally. Hair cell orientations on the utricular epithelium and macula neglecta are similar to those in other bony fishes. Nerve fiber diameters can be divided into three size classes, 1-8 microns, 9-13 microns, and 14 microns or more, with the smallest size class containing the majority of fibers. The distribution of the various classes of fiber diameters is not the same in nerve branches to each of the end organs. Similarly, the ratio of hair cells to axons differs in each end organ. The highest hair cell to axon ratio is in the utricle (23:1) and the smallest is in the macula neglecta (7:1). The number of sensory hair cells far exceed the number of eighth nerve axons in all sensory epithelia.     

Stiffness changes in chick hair bundles following in vitro overstimulation

1. The in vitro effect of intense stimulation on the micromechanical stiffness of hair cell sensory hair bundles was studied at three locations on the chick basilar papilla. Threshold levels of hair bundle motion, produced by a water jet stimulus, were examined before and after exposure to a 300 Hz water jet stimulus set at 25 dB above the pre-exposure threshold level.
2. Threshold levels of motion were systematically examined in 8 unexposed control cells. The level of water jet stimulus needed to achieve the detection threshold of motion remained constant in these cells when periodically tested over a 36.5-min interval.
3. Post-exposure changes in the motion detection threshold of hair bundles were examined in 82 hair bundles, and a number of effects were identified: 2.4% of the hair bundles showed no threshold change; 31.7% of the hair bundles had threshold shifts which indicated an increase in stiffness; 18.3% exhibited a threshold shift that indicated a decrease in hair bundle stiffness, but with no recovery; and 47.6% had thresholds that indicated a decrease in hair bundle stiffness with recovery to pre-exposure levels within 16–18 min.
4. The results suggest that chick hair bundles exhibit complex and varied responses to overstimulation which are very different from that seen in the mammal.

     

Ultrastructure of epidermal sensory receptors in Amphibolurus barbatus (Lacertilis: Agamidae)

Summary The histological and ultrastructural organisation of the epidermal sensory organs in Amphibolurus barbatus has been described with respect to their position and possible functions. The sensory organs, located at the scale's edge, are most numerous in scales of the dorsal surface of the head. Most other scales of the body surface have two receptors located laterally to the spine or keel of the scale. In the imbricate scales of the ventral body region, the receptors lie just beneath the reinforced scale lip. Scanning electron microscopy has revealed the surface of the organ to be a crater lacking any surface projections. These sensory organs have a dermal papilla consisting of a nerve plexus and loose connective tissue. The nerve fibres arising from the plexus, pass to the epidermal columnar cells, where some form nerve terminals at the base of the cells, while others pass between them to form nerve terminals embedded in a superficial layer of cuboidal cells. The superficial terminals are held against the overlying keratin by masses of tonofilaments. The keratin is thickened to form a collar around the periphery of the organ but is only about 0.5 m thick immediately above it. Mechanical deformation of the scale's spine or reinforced scale lip may initiate stimulation of the nerve terminals described.     

Light- and electron-microscopic analysis of a complex sensory organ: The tegula of Locusta migratoria

Summary Structure and organization of the tegula, a cupola-shaped structure located at the anterior base of the wings of locusts, is described using various morphological methods. Based on histological and cytological criteria, two different sensory systems are distinguished: (1) a field of mechanoreceptive hairs, and (2) a chordotonal organ. The total number of sensory cells corresponds to the number of axons within the nerve supporting the tegula. The hairs are situated at the posterior region of the tegula, and each hair is innervated by only one sensory cell. The complex architecture of the chordotonal organ is analyzed and the attachment of the scolopidia to the cuticle is described. A single scolopidium makes contact with several epidermal cells. The attachment cells run in parallel and are oriented longitudinally within the tegula, being connected to each other and to the epidermal cells by desmosomes. A function in relation to wing movements during flight is suggested for the two sensory systems within the mixed sense organ, tegula.     

Comparative features of the surface morphology of the basilar papilla in five families of salamanders (Amphibia; Caudata)

The surface morphology of the basilar recess and papilla was examined in 14 species of newts and salamanders selected from the five families of urodeles (Ambystomatidae, Salamandridae, Hynobiidae, Cryptobranchidae, and Amphiumidae) known to have this end-organ. In this sampling, the general organization of basilar structures is essentially similar across species investigated. The recess forms a tubular diverticulum of the proximal part of the lagena. One wall of the recess is associated with a diverticulum of the intracapsular periotic sac, and an adjacent wall is occupied by the basilar papilla. The papilla contained from as few as five hair cells in specimens of Taricha torosa to over 200 hair cells in Cryptobranchus allegheniensis. In most species, the papilla showed a morphological continuum between tall centrally or distally placed ciliary bundles and short ciliary bundles near the papillar margins. In certain species examined, tall bundles had kinocilia with swellings near their tips. Most forms showed a tendency to have groups of ciliary bundles morphologically polarized either toward or away from the saccule. In Cryptobranchus and Dicamptodon, many bundles had a random orientation. The gross and fine structural features of the basilar complex are compared in urodeles and anurans, and "generalized" features for the amphibian basilar complex are suggested. The basilar complex of Cryptobranchus is interpreted as being most generalized, representing a structural form from which most features of the basilar complex in other urodeles and anurans can be derived.     

Ultrastructure of the osphradium of Aplysia californica

Summary The osphradium of Aplysia californica, a sensory organ, is a small yellow-brown epithelial patch located in the mantle cavity immediately anterior to the rostral attachment of the gill. Scanning electron microscopy reveals a round ellipsoid structure of 0.6–1 mm in diameter with a central, occasionally folded, sensory epithelium. The central area is covered with microvilli and surrounded by a densely ciliated epithelium. Transmission electron micrographs show that the columnar supporting cells in the sensory epithelium contain an abundance of apical pigment granules and microvilli. Between the epithelial-supporting cells, the putative sensory elements consist of thin neurites (0.4–1.5 m in diameter) that reach the sea-water side of the osphradium. The neurites contain many neurotubules, mitochondria, vesicles and cilia in their apices. The nerve endings originate from cell bodies up to 40 m below the epithelium or in the osphradial ganglion itself, as revealed by electron microscopy and retrograde labeling with Lucifer yellow. There appear to be two populations of putative sensory cells, a large population of heavily stained cell bodies 4–10 m in diameter and a few scattered cells of large diameter (25–60 m). Following lanthanum impregnation, septate junctions can be seen between all types of cells in the epithelium, 3–5 m below the sea-water surface. This study provides new information for further investigation of osmo- and mechanosensation in Aplysia californica.     

A Qualitative and Quantitative Study of a Sensory Epithelium in the Inner Ear of a Fish (Colisa labiosa; Anabantidae)

The macula lagenae of the anabantide fish Colisa labiosa was studied with light and transmission electron microscopy. (1) The sensory area is naturally divided in a central area (A) surrounded by a peripheral part (B). (2) Generally the central hair cells are separated by supporting cells, while the peripheral hair cells are found in groups. The cells of a group are not separated by supporting cells. (3) Tubuli-like structures, hexagonal in cross section, are found in all cells. In peripheral hair cells the longitudinally oriented tubuli-like structures are aggregated in thick bundles. (4) Variation in shape, electron density, stereocilia arrangement and size of mitochondria was found in different hair cells. (5) The central hair cells contain large accumulations of presynaptic bodies (10–44). Contrarily, the peripheral hair cells contain only a few pre-synaptic bodies (1–3). (6) The central hair cells are innervated by thick afferent (6–15 μm) and fine presumed efferent (less than 1 μm nerve fibres, while the peripheral hair cells are innervated by thin (1–6 μm) afferent nerve fibres only.     

Central nervous projections of mechanoreceptors in the spider Cupiennius salei Keys.

Summary The basic organization of sensory projections in the suboesophageal central nervous system of a spider (Cupiennius salei Keys.) was analyzed with anterograde cobalt fills and a modified Golgi rapid method. The projections of three lyriform slit sense organs and of tactile hairs located proximally on the legs are described and related to central nerve tracts. There are five main longitudinal sensory tracts in the central region of the suboesophageal nervous mass arranged one above the other. Whereas the three dorsal ones contain fibers from the lyriform organs, the two ventral ones contain axons from the hair receptors. Axons from all three lyriform organs have typical shapes and widely arborizing ipsilateral intersegmental branches and a few contralateral ones. The terminal branches of the afferent projections from identical lyriform organs on each leg form characteristic longitudinal pathways, typical of each organ: U-shaped, O-shaped, or two parallel bundles. The terminations of the hair sensilla are ipsilateral and intersegmental. Two large bilaterally arranged longitudinal sensory association tracts receive inputs from all legs including the dense arborizations from tactile hairs, lyriform organs, and other sense organs. These tracts may serve as important integrating neuropils of the suboesophageal central nervous system.     

Chiton integument: Ultrastructure of the sensory hairs of Mopalia muscosa (Mollusca: Polyplacophora)

Summary The dorsal integument of the girdle of the chiton Mopalia muscosa is covered by a chitinous cuticle about 0.1 mm in thickness. Within the cuticle are fusiform spicules composed of a central mass of pigment granules surrounded by a layer of calcium carbonate crystals. Tapered, curved chitinous hairs with a groove on the mesial surface pass through the cuticle and protrude above the surface. The spicules are produced by specialized groups of epidermal cells called spiniferous papillae and the hairs are produced by trichogenous papillae. Processes of pigment cells containing green granules are scattered among the cells of each type of papilla and among the common epidermal cells.The wall or cortex of each hair is composed of two layers. The cortex surrounds a central medulla that contains matrix material of low density and from 1 to 20 axial bundles of dendrites. The number of bundles within the medulla varies with the size of the hair. Each bundle contains from 1 to 25 dendrites ensheathed by processes of supporting cells. The dendrites and supporting sheath arise from epidermal cells of the central part of the papilla. At the base of each trichogenous papilla are several nerves that pass into the dermis. Two questions remain unresolved. The function of the hairs is unknown, and we have not determined whether the sensory cells are primary sensory neurons or secondary sensory cells.     

Transmissions- und rasterelektronen-mikroskopische Untersuchung an den Sinnesknospen der Tentakeln von Myxine glutinosa L. (Cyclostomata)

Sensory buds of different sizes have been found in the epidermis from the barbels of Myxine glutinosa. The electron microscopic investigations reveal their composition as one type of receptor cells and two types of supporting cells. The receptor cells have apical stereocilia with filamentous internal structure and there are microvilli on the supporting cells. The cytoplasma of the receptor cells contain fibrils and fibril bundles. The sensory buds are innervated by a nerve plexus below the basal lamina with nerve endings between the basal cells, which lie below the sensory buds. A comparison with sensory buds in some other vertebrates is drawn.     

The ultrastructure of taste and touch receptors of the Frog's taste organ

Summary The taste buds from fungiform papillae and the hard palate of frogs were investigated with the scanning and transmission electron microscopes. An immature pre-taste cell and a mature taste cell can be differentiated. Only the mature taste cell exhibits synaptic contact with the afferent taste fibre. Glandular and satellite supporting cells envelop the thin apical processes of the sensory cells. At the base of the taste disc up to 10 Merkel cells form a complex with nerve endings. There are two types of myelinated fibres, large and small. The small fibre innervates the taste cells, the thicker nerve fibre the Merkel cells. The occurrence of two types of receptors explains physiological results.Supported by the Deutsche Forschungsgemeinschaft Rezeptorphysiologie.     

Electron Microscope Observations on in Vitro Cultures of the Isolated Fowl Embryo Otocyst

In vitro cultures of isolated fowl embryo otocysts were studied with the electron microscope. Hair cells of the developing organ of Corti and crista ampullaris have been examined with particular reference to the structure of the cilia and of the cell membrane. Two types of hair cells could be distinguished on the basis whether or not they possessed a "kinocilium" and "stereocilia," or "stereocilia" only. The cytoplasmic membranes were simple and there were no multiple vesicular layers in any of the hair cells. The supporting elements consisted of supporting cells flanking the hair cells, fibroblasts, and the cartilaginous otic capsule. Both the cochlear and vestibular sensory area showed rich innervation by mainly non-myelinated fibers with partial myelinization in others. There were well developed ganglion cells present. Bare axons penetrated the basement membrane and spread, amongst the supporting cells sheltering them, to the base of the hair cells where they formed bud-shaped nerve endings but, at the stage of development examined, no calyces. These in vitro cultures of the isolated fowl embryo otocyst provided convenient and suitable material for the electron microscope study of the sensory epithelium of the ear and revealed further that the isolated fowl embryo otocyst possesses great powers of self-differentiation also at the ultrastructural level.     

Functional differentiation of sensory cells in the avian auditory periphery

Summary Mammals and birds have independently developed different populations of sensory cells grouped across the width of their auditory papillae. Although in mammals there is clear evidence for disparate functions for the two hair-cell populations, the different anatomical pattern in birds has made comparisons difficult. In two species of birds, we have used single-fibre staining techniques to trace physiologically-characterized primary auditory nerve fibres to their peripheral synapses. As in mammals, acoustically-active afferent fibres of these birds innervate exclusively the neurally-lying group of hair cells in a 11 relationship, suggesting important parallels in the functional organization of the auditory papillae in these two vertebrate classes. In addition, we found a strong trend of the threshold to acoustic stimuli at the characteristic frequency across the width of the avian papilla.Abbreviations IHC inner hair cell(s) - OHC outer hair cell(s) - SHC short hair cell(s) - THC tall hair cell(s)     

Morphology of the basilar papilla of the budgerigar,Melopsittacus undulatus

The budgerigar is a representative of the parrot-like birds that, like song birds, have developed complex communication signals. This species is interesting in a psychoacoustic sense, in that it shows unusually good frequency discriminative abilities above about 1 kHz. To begin to understand whether the peripheral hearing organ plays a role in such specializations, we have carried out a quantitative study of the fine anatomy of the basilar papilla and compared it to data from other avian species. The budgerigar basilar papilla is about 2.5 mm long in the living animal and contains about 5,400 hair cells. The hair cells of the papilla show regional specializations similar to those found in other birds and are described from scanning electron microscopic and light microscopic studies. Regiona changes in the basilar papilla, and in the basilar and tectorial membranes are described from light microscopic data. As noted for other avian species, the constellation of morphologic features found in the budgerigar is unique. In general, the hair cell patterns of the budgerigar papilla showed fewer specializations than found in, e.g., a songbird, the starling, but more than seen in a primitive land bird, e.g., the pigeon. There were no features that were obviously related to the unusal psychoacoustic performance of this species. © 1993 Wiley-Liss, Inc.     

The innervation of the myometrium of the cynomolgus monkey (Macaca fascicularis)

Summary The autonomic innervation of the myometrium of Macaca fascicularis consists of bundles of unmyelinated nerve fibres running between the smooth muscle cells, and is therefore considered to be of the fascicular (= unitary) type. Close contacts between nerve fibres and smooth muscle fibres were not found. Modification of the chromaffin method according to Tranzer and Richards made it possible to visualize the heterogeneity of the nerve fibres in a single bundle. The following fibre types were found to coexist: (1) noradrenergic fibres containing synaptic vesicles with a dense granule, (2) cholinergic fibres containing empty synaptic vesicles, and (3) non-adrenergic noncholinergic (NANC) fibres containing only or predominantly large dense-cored vesicles, which do not react with this method. Noradrenergic fibres are the most numerous (around 60%), followed by NANC fibres (30%) and cholinergic elements (around 10%). The distribution of these three types is similar in the cervix, the isthmus and the body of the uterus in pregnant and non-pregnant females.     

Upstream non-coding region which confers polar expression to Ri plasmid root inducing gene rolB

Summary Root differentiation could be elicited on carrot discs by transformation with the agropine Ri plasmid rolB gene cloned in the binary vector Bin19, provided two conditions were met. Firstly, an adequate auxin supply had to be provided. This was achieved by co-inoculation with a strain carrying only the auxin synthetic genes of the T_R-DNA. Most of the resulting roots were then shown to harbour only rolB and no aux genes. Secondly, an extended non-coding region (1200 bp) at the 5 end of rolB had to be included in the construction. A shorter (300 bp) 5 region, including TATA and CCAAT boxes, was not sufficient to trigger root differentiation. Both the extended (B1185) and reduced (B310) 5 regions of rolB were then cloned upstream of the -glucuronidase (GUS) reporter gene and infections carried out both on the apical and on the basal side of carrot discs. Strong expression of GUS, visualized histochemically as an intense blue colouring of transformed cells was observed with B1185-GUS constructions on the apical side of the discs. Only occasionally could coloured cells be observed on the basal side of the discs with B1185-GUS and on both apical and basal sides with B310-GUS constructions. Strong GUS expression was, on the contrary, achieved on cells of both auxin-rich (apical) and auxin-depleted (basal) sides of the discs with the strong constitutive viral promoter, CaMV35S. These results indicate the presence of an upstream regulatory region which confers polar expression to the rolB gene and suggest a role for auxin in its activation.