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Surface ultrastructure of the gill arch of the killifish, Fundulus heteroclitus, from seawater and freshwater, with special reference to the morphology of apical crypts of chloride cells.

The surface ultrastructure of the gill arches of the killifish, Fundulus heteroclitus, adapted to seawater or freshwater, was found to be similar to that reported for other euryhaline teleosts. Two rows of gill filaments (about 42 filaments per row) extended posterolaterally, and two rows of gill rakers (about 10 rakers per row) extended anteromedially from each arch. Leaf-like respiratory lamellae protruded along both sides of each filament, from its base to its apex. The distributions, sizes, and numbers of various surface cells and structures were also determined. All surfaces were covered by a mosaic of pavement cells, which measured about 7 X 4 microns and exhibited concentrically arranged surface ridges. Taste buds were especially prominent on the rakers and the pharyngeal surfaces of the first and second gill arches, but were often replaced by horny spines on the third and fourth gill arches. Apical crypts of chloride cells occurred mostly on the surfaces of the gill filaments adjacent to the afferent artery of the filament. In seawater adapted killifish, crypts resembled narrow, deep holes along the borders of adjacent pavement cells, had openings of about 2 microns2, and occurred at a frequency of about 1 per 70 microns2 of surface area. In freshwater fish, the crypts usually had larger openings (about 10 microns2), occurred less frequently (1 per 123 microns2), and exhibited many cellular projections in their interiors. Changes in crypt morphology may be related to the ion transport function of chloride cells.

Adaptation, Physiological↗

Relation between PROP (6-n-propylthiouracil) taster status, taste anatomy and dietary intake measures for young men and women.

Previous studies have related 6-n-propylthiouracil (PROP) taster status to preference for, and consumption of various (bitter-tasting) foods recognized for their cancer-preventive properties. The aim of this study was to examine PROP taster status in relation to general measures of dietary intake as well as the consumption of specific food groups. College students (n=183) were classified as non-tasters (n=49), medium tasters (n=89) and supertasters (n=45) of PROP based on intensity ratings of NaCl and PROP solutions. Dietary intake measures were derived from a food frequency questionnaire and body mass indices (BMI) were derived from self-reported height and weight. Supertasters had higher fungiform papillae counts on the anterior tongue than tasters and non-tasters, yet the distributions of papillae counts overlapped across PROP taster groups. No significant differences were found for BMI values and energy intake among taster groups. PROP-tasting women derived a greater percentage of their dietary energy from fat, and consumed less fruit than non-tasters. PROP supertasters did not differ from tasters and non-tasters in intake of bitter fruits, vegetables or beverages except for a lesser intake of green salad. The hypothesis that PROP supertasters, through heightened sensitivity to, and avoidance of, bitter-tasting fruits, vegetables and other foods with antioxidant properties, may therefore be at increased risk for diet-linked diseases such as cancer, is not supported by these findings.

Adolescent↗

Gustin from human parotid saliva is carbonic anhydrase VI.

Gustin, a zinc-metalloprotein constituting about 3% of human parotid saliva protein was previously isolated and characterized as a single polypeptide chain of 37kDa with one mole of zinc tightly bound to the protein. It exhibited biological activity activating calmodulin dependent bovine brain cAMP phosphodiesterase and was decreased in saliva of patients with loss of taste in whom taste buds showed a specific pathological morphology. Determination of its primary structure by amino acid sequence revealed it was identical with carbonic anhydrase (CA) [EC 4.2.1.1] VI and had two N-linked glycosylation sites. Analysis by reverse phase HPLC and SDS-PAGE before and after deglycosylation confirmed a single peak with molecular weight of the purified protein being 37kDa, the deglycosylated protein, 33kDa. N-linked carbohydrate chains contained N-acetyl glucosamine, galactose, mannose, and fucose interior to di, tri and tetra sialyated termini. By isoelectric focusing five increasingly acidic pI values were determined consistent with addition of sialic acid as the terminal carbohydrate residue on the N-linked glycoforms of the protein. Gustin was found to exhibit CA activity but was inhibited by known CA inhibitors in a different manner than CA I or II. These findings, consistent with analysis of previous investigators, indicate that parotid saliva gustin is CA VI.

Amino Acid Sequence↗

Identification of coding single-nucleotide polymorphisms in human taste receptor genes involving bitter tasting.

T2Rs comprise a G-protein-coupled receptor superfamily that contains functionally defined bitter taste receptors. Here we report the tissue expressions and coding single-nucleotide polymorphisms (cSNPs) in human T2R genes (hT2R3, hT2R4, and hT2R5) on chromosome 7q31. We first demonstrated that hT2R3, hT2R4, and hT2R5 are actually expressed in the circumvallate papillae of the human tongue by reverse transcription-polymerase chain reaction (RT-PCR). We identified six cSNPs within the T2R receptor genes. The hT2R4 and hT2R5 contained four and one cSNPs that cause missense mutations, respectively, while hT2R3 included one silent nucleotide mutation. However, we could not find any nonsense mutations that resulted in a frameshift or a premature stop codon within the open reading frames. Genotype frequencies of each cSNP were in Hardy-Weinberg equilibrium. The identification of nucleotide diversity and amino acid polymorphisms in human T2R receptors could help clarify individual differences in the acceptability and sensitivity to bitter compounds.

Base Sequence↗

Acute effects evaluation of HgCl2 on epidermis of Trichomycterus brasiliensis (Siluroidei; trichomycteridae).

The effects of HgCl2 on the epidermis of Trichomycterus brasiliensis were studied by histological, histochemical, and ultrastructural methods. First, the normal organization of the tissue was described in order to study the HgCl2 effects on the skin structure. The epithelial cells presented a typical structural organization found in many fishes. Basically five types of cells could be detailed: goblet cells, club cells, germinative stratum cells, intermediate layer cells, and epithelial cells in the superficial layer. The goblet cells in the superficial layer present evidence of secretory activity by positive histochemical reactions. In cells exposed to HgCl2, different morphological alterations were observed in the epithelium structure, such as an increase in the lymphocyte number, hypertrophied epithelial cells at the surface, modified taste buds, obstruction of the goblet cells pore, and high cellular proliferation. Moreover, the chemical nature of the goblet cells was not modified by the presence of the HgCl2 dissolved in water. All animals died within 24 hr after the contamination with inorganic mercury at concentrations of 0.2 and 0.1 mg HgCl2/liter.

Animals↗

High-voltage electron microscopy and 3-D reconstruction of solitary chemosensory cells in the anterior dorsal fin of the Gadid fish Ciliata mustela (Teleostei).

Solitary chemosensory cells (SCCs) are secondary sensory cells present in the epidermis of most primary aquatic vertebrates. In rocklings, the epidermis of the anterior dorsal fin (ADF) contains approximately 5 million SCCs. High-voltage electron microscopy and three-dimensional reconstructions from serial sections were used to examine the ultrastructure, arrangement, and synaptic contacts of the SCCs in the rockling ADF. Approximately 15% of all cells in the fin ray epidermis are SCCs, which occupy roughly 30% of the epidermal volume. These spindle-shaped cells are 25-30 microm long and up to 10 microm wide and terminate apically in a microvillus protruding 2-5 microm above the epidermal surface. SCCs contain abundant endoplasmic reticulum and a large Golgi apparatus in their proximal regions. The distal parts of SCCs contain characteristic vesicles, elongate mitochondria, and longitudinal strands of intermediate filaments. Synapses between SCCs and nerves resemble those found in teleost taste buds. One to four synaptic contacts per SCC were found. We hypothesize that the apparent secretory activity of the SCCs serves to replenish the apical membrane and mucus. Furthermore, parallel sampling of several hundred SCCs by single nerve fibers may serve low-threshold detection rather than stimulus localization.

Animals↗

Distribution of calcitonin gene-related peptide nerve fibers in the canine larynx.

Immunohistochemistry was used to investigate the distribution pattern of calcitonin gene-related peptide (CGRP) nerve fibers in the laryngeal mucosa, glands and intrinsic muscles of the dog. CGRP immunoreactive nerve fibers were found more frequently than substance P immunoreactive nerve fibers in every region of the larynx. In the epithelia, CGRP nerve fibers were mainly found in the epiglottis, arytenoid region and subglottis. Many taste buds were observed in the arytenoid region and were densely innervated by the CGRP nerve fibers. In the lamina propria, the plexus of CGRP nerve fibers was present, with some of these fibers associated with blood vessels. Laryngeal glands were also innervated by a few CGRP nerve fibers. In the intrinsic laryngeal muscles, abundant immunoreactivity was observed and many motor end-plate-like structures were found with CGRP immunoreactivity. These findings strongly suggest that CGRP plays an important role in all of the sensory, motor and autonomic nervous systems of the larynx.

Animals↗

Tissue expression of lipocalins in human lacrimal and von Ebner's glands: colocalization with lysozyme.

BACKGROUND: Tear-specific prealbumin is a group of proteins recently renamed as the tear lipocalins. These proteins were initially described as unique to lacrimal fluid. The tissue distribution and localization have never been thoroughly studied. METHODS: The distribution of purified tear lipocalins was studied in many human secretions and tissues by western blots, immunohistochemistry and immunoelectron microscopy. RESULTS: Tear lipocalin species of the same molecular weights were observed in western blot lanes loaded with tears, saliva, and protein extracts from the lacrimal and lingual von Ebner's glands. Lacrimal and von Ebner's glands contained tear lipocalins; other human tissues and secretions, including other salivary glands and taste buds, did not. Tear lipocalins colocalized with lysozyme in serous acinar cells of lacrimal and von Ebner's glands. Ultrastructurally, tear lipocalins were present on polyribosomes, endoplasmic reticulum, and Golgi areas. Lipocalins were concentrated in lacrimal secretory granules in amounts commensurate with a regulated pathway. CONCLUSION: Tear lipocalins are expressed and truncated similarly in lingual von Ebner's and lacrimal glands, but not at all in other human tissues. Lipocalins are expressed and secreted with lysozyme. Lipocalins are concentrated in secretory granules in an amount consistent with a regulated secretory pathway.

Amino Acid Sequence↗

Stereo architecture of the connective tissue cores of the lingual papillae in the treeshrew (Tupaia glis).

The stereo architecture of the lingual connective tissue cores (CTC) in the treeshrew (Tupaia glis) (which has the primitive characteristics of primates) was observed by scanning electron microscopy, and compared to that of other animal orders. The tongue of the treeshrew has three vallate papillae which are situated in the posterior part of the tongue, while some macaques have several vallate papillae. Among numerous filiform papillae, fungiform papillae are sporadically distributed. A filiform papilla consists of a bundle of several slender spine-like processes arranged in a circle at the basal margin. After removal of the epithelium, the CTC of the filiform papilla looks like a human hand raised with the palm facing towards the tongue tip. The fungiform CTC in the threeshrew is columnar in shape (rather similar to that of Insectivora and Rodentia) and at the top there are several round depressions for taste buds. In the treeshrew several large rod-shaped processes are derived from the postero-lateral margin of the tongue, as in Carnivora (dogs and cats), where foliate papillae are located in many other animal species. The treeshrew has numerous characteristics similar to those of the crab-eating macaque (Primates), but at the same time it has some characteristics similar to those of Insectivora, Rodentia, Carnivora and Artiodactyla.

Animals↗

The development of the eye in Astyanax mexicanus (Characidae, Pisces), its blind cave derivative, Anoptichthys jordani (Characidae, Pisces), and their crossbreds. A scanning- and transmission-electron microscopic study.

The development of the eye of the characin Astyanax mexicanus, of its blind derivative Anoptichthys jordani, and crossbreds of both forms was studied at different ontogenetic stages by means of scanning- and transmission-electron microscopy. Astyanax exhibits a form of eye development resembling that in other characid species. A severe reduction of the eye could be observed in Anoptichthys starting with the second day of ontogeny. This degenerational process is characterized by the following features: 1) An overgrowth of epidermal tissue that gradually covers the surface of the eyeball; 2) the sinking of the eyeball below the surface of the integument; 3) the formation of epidermal channels from the body surface to the disappearing surface of the eyeball; 4) a severe degeneration of the retinal sensory cells; and 5) a small number of pigment granules in the pigment epithelial cells. The progeny of crosses between Astyanax and Anoptichthys show varying degrees of these degenerational signs. Taste buds and the lateral line organ display identical features in all crosses analyzed with the scanning electron microscope.

Animals↗

On the retinal pigment epithelium of the barn owl (Tyto alba).

The fine structure of the retinal pigment epithelium (RPE) of the barn owl (Tyto alba) is described. In general it resembles that of the retinal pigment epithelium of other vertebrate animals and a high content of myeloid bodies, lipid droplets, electron dense cytosomes, and especially phagosomes is observed. The latter show different structural specializations which include membranes, tubules, filaments, granules, vesicles, and crystalloid patterns. The central part of the retina often contains a peculiar inclusion body which is separated from the cytoplasm by one or two distinct membranes and is composed of a regularly arranged tubular network which forms a three-dimensional honeycomb structure. The tubules in part are in continuation with randomly oriented and densly packed tubules and stacks of discs consisting of paired lamellae. Similar formations within secretory cells of a gland (dendritic organ) and the taste buds of catfish are discussed.

Animals↗

A gustatory mutant of Drosophila defective in pyranose receptors.

Mutations in an X-linked gene, gust-A, block the responses of Drosophila melanogaster to a group of pyranose sugars. It is shown that the behavioural effects of this mutation are correlated with a loss of electrical responses in taste receptors. The mutation affects the chemoacceptors for pyranose sugars leaving the furanose acceptors intact.

Animals↗

[Evaluating function and disorders of taste].

In a first anatomical section the peripheral gustatory pathways, their central connections, nuclei and cortical projections are discussed. It is evident, that the gustatory fibres from the posterior part of the tongue run in the IX nerve and those from the soft palate reach the medulla oblongata via the petrosal and facial nerve. For the anterior part of the tongue there obviously exists only one gustatory pathway via the chorda tympani-facial nerve. About the further central pathways of taste fibres is much less known. In a second part the methods of taste testing with different taste solutions and the electrogustometry are described. Their practical use and the pitfalls of testing are considered. The disorders of the taste sense compose a third part. Genetic and endocrine abnormalities as well as the side effects of drugs and radiotherapy and the destruction of taste nerves may lead to gustatory deviations. The possibility of central gustatory disorders, especially the combined loss of taste and smell as a result of trauma are mentioned. A chapter dealing with the therapy of taste disorders and a short outlook on the genetic aspects of this oral sense complete this review.

Afferent Pathways↗

Bitter taste receptors and human bitter taste perception.

A vast number of structurally diverse bitter compounds need to be detected by a subfamily of only approximately 25 human bitter receptors. Failure in detecting them might be lethal, since some naturally occurring bitter compounds, such as strychnine, are very toxic. This review presents an overview about the enormous progress in the field of mammalian bitter taste research with special emphasis on humans, if data were available. It summarizes the current knowledge about the anatomical basis for bitter taste perception, intracellular signal transduction, evolution, expression and polymorphisms of hTAS2R genes, and the molecular basis for the recognition of bitter compounds.

Animals↗

Three sweet receptor genes are clustered in human chromosome 1.

A search of the human genome database led us to identify three human candidate taste receptors, hT1R1, hT1R2, and hT1R3, which contain seven transmembrane domains. All three genes map to a small region of Chromosome (Chr) 1. This region is syntenous to the distal end of Chr 4 in mouse, which contains the Sac (saccharin preference) locus that is involved in detecting sweet tastants. A genetic marker, DVL1, which is linked to the Sac locus, is within 1700 bp of human T1R3. Recently, the murine T1Rs and its human ortholog have been independently identified in combination as sweet and umami receptors near the Sac locus. All three hT1Rs genes are expressed selectively in human taste receptor cells in the fungiform papillae, consistent with their role in taste perception.

Alleles↗

The apical membrane of intestinal brush cells possesses a specialised, but species-specific, composition of glycoconjugates--on-section and in vivo lectin labelling in rats, guinea-pigs and mice.

Brush cells are specialised epithelial cells that are assumed to represent chemoreceptors of the digestive tract. They comprise a small population of the epithelial cells lining the intestine, possess a unique ultrastructure and, in many aspects, resemble the receptor cells of taste buds. To characterise glycoconjugates possibly involved in a sensory function, we investigated brush cells in the small intestine of three species using lectin histochemistry in confocal light and thin-section electron microscopy. Brush cells of rats were selectively labelled by the sialic acid-specific lectin Maackia amurensis agglutinin, those of guinea-pigs by the D-galactose-specific lectin Bandeiraea simplicifolia agglutinin, isolectin B4 and those of mice by the L-fucose-specific lectin Ulex europaeus agglutinin lectin I. Lectin binding sites were consistently located in the glycocalyx of the apical membrane and in that of cytoplasmic vesicles. In vivo lectin labelling revealed that the glycoconjugates of the apical membrane are accessible under physiological conditions, that brush cells do not endocytose and that they probably possess a high membrane turnover rate. The results show that specialisations exist in the composition of glycoconjugates forming the glycocalyx of brush cells in all species investigated. The presence of brush cell-specific glycoconjugates would be in accordance with the current hypothesis of a receptive function of brush cells. Differences in the specific glycosylation patterns among rats, guinea-pigs and mice indicate that species-specific adaptations exist.

Animals↗

Comparative morphological study of the lingual papillae and their connective tissue cores of the koala.

The stereo structure of each lingual papilla of the koala has a similar structure to that of various other animal species: the koala has a lingual prominence (intermolar prominence) with larger filiform papillae. (A lingual prominence is a characteristic in herbivorous animals.) The external form and connective tissue core (CTC) of the filiform papillae of koalas consist of one large main process and several smaller accessory processes. (These are similar to carnivorous animals.) Fungiform CTC have a thick dome-like structure, with several taste buds on the top. There are three vallate papillae: one central midline and two laterally located vallate papillae. The central vallate papilla has a posterior pouch lined with ciliated and non-ciliated epithelial cells. Long conical papillae are distributed in the posterior lateral area where foliate papillae are distributed in many other animal species. (Finger-like papillae are seen in dog and cat instead of foliate papillae.) It may be suggested that the tongue of the koala evolved in a special environment in Australia. Even though it has still retained special features similar to those of carnivorous cats and dogs it has evolved to resemble the tongues of herbivorous animals.

Animals↗

Artemin-like immunoreactivity in the zebrafish, Danio rerio.

Artemin is a member of the glial cell line-derived neurotrophic factor (GDNF) family. It is a neurotrophic factor that supports neurite migration and outgrowth and survival of the sympathetic and sensory nervous system. Artemin has been studied in human and murine tissues, but no study has been devoted to nonmammalian species. Zebrafish is a teleost fish belonging to the family Ciprinidae, which is becoming an important model species for genetic and developmental studies. Thus, the aim of the present investigation was to evaluate, by immunochemical and immunocytochemical analyses, the tissue distribution pattern of artemin in zebrafish. Different isoforms of artemin with corresponding different molecular weights were detected in the brain, muscle, testis, ovary, kidney, gut, and gills of zebrafish by Western blot analysis. Immunocytochemical analysis showed artemin-like immunoreactivity in different cell types: in glial cells and rare neurons of the central nervous system, taste buds, retina, neuromasts of the lateral line, dorsal root ganglia, sympathetic ganglia, gill epithelium, tubular kidney epithelium, gut epithelium and ganglia, pancreas, thyroid, hypothalamus, testis, and ovary. These results indicate a wide distribution of artemin-like immunoreactivity in adult zebrafish, related to the presence of different forms of artemin. These findings might suggest a complex maturation pattern of artemin, whose forms could also exert different roles in zebrafish tissues.

Animals↗