Seasonal changes in the epidermis of the brown trout Salmo trutta (L.)

A sexual dimorphism was found in the epidermis of hatchery-reared brown trout. The epidermis of both male and female fish undergoes rhythmical changes in thickness during successive spawning cycles. For most of the year the male has a significantly thicker epidermis than the female. During the spawning period (December and January) there is a highly significant drop in the concentration of mucous cells in the epidermis of the male brown trout. These findings are discussed in the light of our knowledge of the skin structure in other salmonid fish and in relation to possible hormonal factors influencing the epidermis of teleosts.     

Observations on the structure and histochemistry of the skin of Mystus gulio (Ham.) (Osteichthyes: Siluriformes, Bagridae)

The skin of Mystus gulio (Ham.) is typical of siluroids in not having any scales. Large number of mucous cells and sacciform granular cells, interspersed with taste buds, were found in the epidermis. Histochemical tests showed the presence of weakly sulphated mucopolysaccharides in the mucous cells, while the sacciform granular club cells were found to be proteinaceous, with a small amount of lipids and phospholipids. The dermis is composed of a stratum laxum and a stratum compactum. The subcutis was found to consist of loosely arranged collagen fibres and fat cells.     

Histochemistry of glycoconjugates in mucous cells of Salmo trutta uninfected and naturally parasitized with intestinal helminths

Mucus secreted onto the surface of the intestine forms a physical barrier to invading parasites so that a possible attachment of helminths to the surface is prevented and their expulsion by peristalsis facilitated. In mammals, intestinal parasites induce hyperplasia and hypertrophy of intestine goblet cells and provoke changes in the mucus composition. In fish, this topic has received less attention. In the present investigation, histochemical methods were employed to compose intestinal mucous cell numbers and their glycoconjugate composition were compared by uninfected brown trout Salmo trutta and in S. trutta parasitized with Cyathocephalus truncatus or Pomphorhynchus laevis. When P. laevis was present in the intestine of the brown trout, the total mucous cell number, and the number of mucous cells containing acid or mixed glycoconjugates were significantly enhanced. No significant change in the total mucous cell number was detected in the intestine of fish parasitized with C. truncatus in comparison with uninfected brown trout. A significant increase was observed in the number of both acid (especially sulphated) and mixed glycoconjugates containing mucous cells as well as a significant decrease in the number of neutral glycoconjugates containing mucous cells. When intestinal helminths were present, the thickness of the adherent mucous gel increased. In a limited number of other fish species, the occurrence of gill and intestinal parasites has been reported to increase the mucosal glycoconjugate secretions. Our study is the first quantitative report on the effects of intestinal helminths on the density of mucous cells and mucus composition in a fish species.     

Acidophilic granular cells in the epidermis of the brown trout,Salmo trutta L.

Summary Acidophilic cells occur in the epidermis of several species of salmonid fish, although their abundance fluctuates considerably between individuals within the same population and at different times during the life cycle. The histology, histochemistry and ultrastructure of an acidophilic, granular celltype in the epidermis of the brown trout, Salmo trutta L., is described. At the light microscope level this cell type is easily distinguished from the large, mucus-secreting, epidermal goblet cells by its acidophilic, proteinaceous secretion. At the ultrastructural level this secretion consists of membrane-bound granules formed by the very active Golgi region. It is argued that the acidophilic, granular cell is not a transformed blood cell but constitutes a normal epidermal component of the brown trout. Possible roles of this cell in the function(s) of the epidermis are discussed.     

D-Glucose uptake in fish hepatocytes: mediated by transporter in rainbow trout (Oncorhynchus mykiss), but only by diffusion in prespawning lamprey (Lampetra fluviatilis) and in RTH-149 cell line

The transport of D-glucose into rainbow trout (Oncorhynchus mykiss) and river lamprey (Lampetra fluviatilis) hepatocytes, as well as into rainbow trout hepatoblastoma cell line RTH-149 was studied using tracer methods. The half-time for D-glucose equilibration was 15 s for rainbow trout. The half-times for the non-metabolizable D-glucose analog, 3-O-methyl-D-glucose equilibration were 8 s, 37 s and 38 s for rainbow trout, lamprey and RTH-149 cells, respectively. The 3-O-methyl-D-glucose was taken up by rainbow trout hepatocytes by facilitated diffusion in addition to simple diffusion. The uptake showed saturation kinetics with the K(m) of 37 mM and V(max) of 62 mmol kg(-1) cells min(-1). The uptake was sensitive to phloretin and cytochalasin B, but not affected by ouabain. The 3-O-methyl-D-glucose uptake by lamprey hepatocytes and RTH-149 cells showed no indication of saturation up to 160 mM, and was not affected by phloretin, cytochalasin B or ouabain, which suggests the mode of transport to be by passive diffusion. However, immunocytochemical stainings revealed the existence of mammalian type GLUT1 and GLUT2 transporters in all cells studied. The lack of a functioning carrier-mediated glucose uptake in lamprey hepatocytes might be due to its physiological state (prespawning starvation). The minor 3-O-methyl-D-glucose uptake into RTH-149 cells compared to freshly isolated rainbow trout hepatocytes might reflect low metabolic activity of the cell lines. Under the conditions applied the RTH-149 cell line is no suitable in vitro model for glucose transport in fish cells.     

Vitamin D regulated keratinocyte differentiation

The epidermis is the largest organ in the body. It is comprised primarily of keratinocytes which are arranged in layers that recapitulates their programmed life cycle. Proliferating keratinocytes are on the bottom-the stratum basale. As keratinocytes leave the stratum basale they begin to differentiate, culminating in the enucleated stratum corneum which has the major role of permeability barrier. Calcium and the active metabolite of vitamin D, 1,25(OH)(2)D(3), play important roles in this differentiation process. The epidermis has a gradient of calcium with lowest concentrations in the stratum basale, and highest concentrations in the stratum granulosum where proteins critical for barrier function are produced. Vitamin D is made in different layers of the epidermis, but 1,25(OH)(2)D(3) is made primarily in the stratum basale. Together calcium and 1,25(OH)(2)D(3) regulate the ordered differentiation process by the sequential turning on and off the genes producing the elements required for differentiation as well as activating those enzymes involved in differentiation. Animal models in which the sensing mechanism for calcium, the receptor for 1,25(OH)(2)D(3), or the enzyme producing 1,25(OH)(2)D(3) have been rendered inoperative demonstrate the importance of these mechanisms for the differentiation process, although each animal model has its own phenotype. This review will examine the mechanisms by which calcium and 1,25(OH)(2)D(3) interact to control epidermal differentiation.     

Histochemical studies of mucosubstances in the epidermis of a nonscaly teleost Mystus (Mystus) vittatus Bl. (Pisces-Bagridae)

The epidermis of Mystus (Mystus) vittatus contains two well differentiated mucous cells which secrete different mucosubstances. The goblet cells contain periodate reactive neutral mucosubstances, glycogen, testicular hyaluronidase resistant sulphated mucosubstances, and sialic acid rich glycoproteins. The clavate cells contain small amounts of neutral and sulphated mucosubstances and no glycoproteins. The difference in the histochemical nature of the two types of mucous cells is discussed in relation to their physiological activities.     

Distribution of ferric iron in larval lampreys, Petromyzon marinus L

The distribution and abundance of ferric iron in larval lampreys (Petromyzon marinus L.) were investigated using light microscopy and the Prussian blue stain. Animals from various watersheds contained different concentrations of iron, although the sites of deposition were the same for all animals. A major portion of iron is within adipose tissue, while the liver, and cartilage contain predominantly low to trace amounts of iron, respectively. Iron is associated with fibrous connective tissue in several places in the body, and this association may have particular significance in the inner ear. Iron is also located in cells of the meninges. The presence of iron in the epithelial cells of the posterior intestine may reflect elimination of the metal through the extrusion of iron-loaded cells into the intestinal lumen. Iron within mucous cells of the epidermis, suggest elimination of iron during mucous secretion. Iron-loaded cells of bipolar shape are also present in the epidermis, but are particularly prominent around the branchiopore. Low concentrations of iron are observed within in melanin-containing macrophages (melano-macrophages) in regions of iron absorption, erythrophagocytosis, and haemopoiesis. High levels of iron in the epithelia and lumina of pronephric tubules are concomitant with degeneration of this organ. These data are evidence of the wide spread distribution of iron in lamprey tissues and additional evidence for the potential value of lampreys for the study of iron metabolism in vertebrates.     

Molecular cloning and sequencing of the banded dogfish (Triakis scyllia) interleukin-8 cDNA

The dogfish (Triakis scyllia) interleukin-8 (IL-8) cDNA was isolated from mitogen-stimulated peripheral white blood cells (WBCs) utilising the polymerase chain reaction (PCR). The cDNA sequence showed that the dogfish IL-8 clones contained an open reading frame encoding 101 amino acids. A short 5' untranslated region (UTR) of 70 nucleotides and a long 3' UTR of 893 nucleotides were also present in this 1.2-kb cDNA. Furthermore, the 3' UTR of the mRNA contained the AUUUA sequence that has been implicated in shortening of the half-life of several cytokines and growth factors. The predicted IL-8 peptide had one potential N-linked glycosylation site (Asn-72-Thr-74) that is not conserved in other vertebrates. It also contained four cysteine residues (Cys-34, 36, 61 and 77), which are characteristic of CXC subfamily cytokines and found in all vertebrates, to date. The dogfish IL-8 lacked an ELR motif as found in the lamprey and trout. Comparison of the deduced amino acids showed that the dogfish IL-8 sequence shared 50.5, 41.2, 37.1 and 40.4-45.5% identity with the chicken, lamprey, trout and mammalian IL-8 sequences, respectively.     

Ultrastructural and histochemical study on gills and skin of the Senegal sole, Solea senegalensis

This study was undertaken to identify the normal ultrastructural features of gills and skin of the Senegal sole, Solea senegalensis, for a comparative measure to morphological alterations caused by environmental stressors such as reduced water quality and diseases. In the Senegal sole skin, four morphologically distinct layers were identified: cuticle, epidermis, dermis and hypodermis. The epidermis was composed of stratified epithelium containing three cellular layers: the outermost or mucosa layer, the middle or fusiform layer and the stratum germinativum or the basal layer. In the mucosa, two mucous cell types were differentiated: type A cells containing several round vesicles of different electron density and type B cells containing mucosomes of uniform electron density. Senegal sole have five pairs of gill arches, each containing two rows of well‐developed and compactly organized primary filaments and secondary lamellae. Fingerprint‐like microridges were observed on the surface of epithelial cells. The branchial lamellae epithelium consisted of different cell types: pavement, mucous and chloride. Between the chloride cells and the larger pavement cells, accessory cells were observed. Complexes of tight junctions and desmosomes were frequently observed between adjacent chloride and epithelial cells. Neutral mucosubstances and/or glycoconjugates were observed in the epidermis, dermis and hypodermis of S. senegalensis skin. Proteins rich in different amino acids, such as arginine and cysteine, reacted negatively or weakly positive in the epidermis, dermis and hypodermis. In gills, some mucous cells responded weakly positive to periodic acid‐Schiff (PAS) reaction but were strongly stained with Alcian Blue at pH 0.5, 1 and 2.5. When Alcian Blue pH 2.5–PAS reaction was performed, most mucous cells were stained blue (carboxylated mucins) and some mucocytes stained purple, indicating a combination of neutral and acid mucins. Proteins rich in cysteine‐bound sulphydryl (‐SH‐) and cystine disulphide (‐S‐S‐) groups were strongly detected in branchial and epidermal mucous cells, whereas lysine, tyrosine and arginine containing proteins showed very weak staining in both epidermal and branchial mucous cells. Protein reactions were strongly positive in the pillar cells, except for those rich in tryptophan, whereas the branchial cartilaginous tissue did not show an important reaction. The performed lipid reactions were negative in goblet and chloride cells. It is concluded from this study that ultrastructural and cytohistochemical features of the Senegal sole skin and gills may serve as control structures in both natural and aquaculture systems to monitor or detect environmental stress responses at the histological level.     

Histochemical studies of oxidoreductases, acid carbohydrates and glycoproteins in the epidermis of the electric eel Electrophorus electricus

The activities of various oxidative enzymes and the presence of acid carbohydrates and acid glycoproteins in the epithelial cells and the mucous goblet cells of the epidermis of the Teleost Electrophorus electricus have been determined by means of a series of selected histochemical techniques of light microscopy. The enzyme activities show a distribution pattern confined mainly to basal cell layers and outer cell layers with a comparatively lower gradient in the transitional cell layers. A mixture of sialic acids of both N-acylated and O-acylated types is found in the mucous goblet cells and the functional significance of mucous production is related to the first line of defense against pathogenic colonization. The higher incidence of various oxidoreductases distributed throughout the entire epidermis is correlated with their key role which can play in the processes of cell differentiation and cell multiplication except for those regarding keratin formation which is not produced in the epidermis of most fish.     

Structure, histochemistry and the effect of handling on the mucous cells of the epidermis of the char Salvelinus alpinus (L.)

The structure and histochemical characteristics of the epidermis of the char Salvelinus alpinus (L.) were studied at the light microscope level. Sialic acid-containing glycoproteins appear to form the main components of the mucous or goblet cells. A single incidence of handling can significantly increase the concentration of superficial goblet cells in the epidermis. Repeated handling results in a maximal response after 1 week but this is reduced to control levels after 1 month. These results are interpreted in terms of the structure of the char epidermis.     

Histochemical analysis of glycoproteins in the secretory cells in the epidermis of the head skin of Indian Major Carp,Labeo rohita

A series of histochemical procedures were employed to localise and characterise glycoprotein (GP) classes produced by the epithelial cells, the type A and the type B mucous goblet cells (MGCs) and the club cells in the epidermis of Labeo rohita. The epithelial cells secreted GPs with oxidizable vicinal diols and GPs with sialic acid residues without O-acyl substitution in low concentrations. The type A MGCs and the type B MGCs, in contrast, produced these GPs in high concentrations. Further, these MGCs produced GPs with O-sulphate esters as well. GPs with O-sulphate esters were produced in high concentration by the type A MGCs and in low concentration by the type B MGCs. The club cells produced GPs with oxidizable vicinal diols in trace amounts. Production of more than one type of GPs suggested a basis for functional discrimination in their role in the mucous secretions at the skin surface. This is considered an adaptation to environment inhabited by the fish and is discussed in relation to their role in lubrication, protection and inhibition of the invasion and proliferation of pathogenic micro-organisms.     

Merkel cells and nerve endings in the labial epidermis of a lizard

Summary Examination of the labial epidermis of the lizard Lacerta sicula revealed cells displaying all features of Merkel cells. These cells are located in the stratum basale of epidermal pegs and are arranged in clusters.     

Waste-water management plant effluents cause cellular alterations in the skin of brown trout

To assess the impact of a sewage plant on fish, brown trout Salmo trutta were kept in two cages for 55 days in a moderately polluted river upstream of a sewage plant. In one of the cages, undiluted treated waste water of the sewage plant (WWE) was added at an average concentration of 5%, whereas the other cage received river water (R) only. A high mortality occurred in the WWE group. In comparison to control trout held in tap water, the skin structure and ultrastructure were altered clearly in both groups exposed to river water, including necrosis, apoptosis, decreased number of mucous cells, decrease in epidermal thickness, invasion of leucocytes, extension of melanocytes into the epidermis, being gradually more prominent in the WWE group. The most obvious difference between the two exposed groups was found in structure, size and electron density of the secretory vesicles of the filament cells. This and the observed vacuolation of Golgi saccules are indicative for disturbances in the secretory pathway of the filament cells. Certain toxins were suspected to cause the decompaction of myelin sheaths demonstrated in both groups. Reasons for the rather minor overall differences between the exposed groups are discussed. The extremely high mortality rate in the WWE group supports the importance of reducing the load of pollutants in the effluent of the waste-water management plant.     

Biological features of the skin of the pika, Ochotona rufescens rufescens: histological structure and enzymatic histochemical reactivity

To examine the usefulness of the pika, Ochotona rufescens rufescens, as an experimental animal for skin irritability tests, the histological structure and enzymatic histochemical reactivity of pika skin were investigated. The pika had a hair cycle similar to that of the rabbit. The skin and epidermis of the pika trunk were 1.16mm and 29.5 microns thick, on the average, respectively. Both of them were the thickest in the dorsal region followed by the interscapular area, while they were the thinnest in the abdominal region. In the epidermis of the pika, the strata corneum, granulosum, spinosum and basale were rather clearly distinguished. The cell arrangement in the stratum basale was more compact than that in the rabbit. Dermal mast cells, which are distributed in the stratum reticulare in rabbits and guinea pigs, were distributed in the stratum papillare right beneath the epidermis. The mast cell of the pika in the TEM images had granules of low electron density and with relatively long microvilli and rather large mitochondria. The activities of the enzymes, SDH, MDH, LDH, beta HBDH, alpha GPDH, ALD, G6PDH and GPR, in the hair follicles and sebaceous glands of the pika were similar to those of the rabbit.     

Sacciform cells in the epidermis of the brown trout,Salmo trutta,and the Arctic char,Salvetinus alpinus

Summary Sacciform cells containing an acidophilic, proteinaceous secretion, were identified in the epidermis of the brown trout and Arctic char. This cell type increased in number during the chronic stages of infestation by the ectoparasitic flagellate, Ichthyobodo sp., in immature brown trout, and decreased during sexual maturation in male brown trout and char. It is suggested that the salmonid sacciform cell produces a secretion which protects the fish against infestation or damage by skin parasites.     

Immunohistochemical localization of irisin in skin,eye, and thyroid and pineal glands of the crested porcupine (Hystrix cristata)

Irisin was first identified in muscle cells. We detected irisin immunoreactivity in various organs of the crested porcupine (Hystrix cristata). In the epidermis, irisin immunoreactivity was localized mainly in stratum basale, stratum spinosum and stratum granulosum layers; immunoreactivity was not observed in the stratum corneum. In the dermis, irisin was found in the external and internal root sheath, cortex and medulla of hair follicles, and in sebaceous glands. Irisin immunoreactivity was found in the neural retina and skeletal muscle fibers associated with the eye. The pineal and thyroid glands also exhibited irisin immunoreactivity.     

Histochemistry of the unicellular glands in relation to their physiological significance in the epidermis of Monopterus cuchia (Synbranchiformes, Pisces)

The cytochemical nature of the secretory contents of the unicellular glands—the mucous cells and the sacciform granulated cells in the epidermis of Monopterus cuchia has been described in order to understand their physiological significance.
The mucous cells secrete mainly the acid mucopolysaccharides and mucoproteins in addition to small amounts of phospholipids. The secretory contents of the sacciform granulated cells are proteinaceous in nature and give strong reactions for elastin, tryptophan and tryosine. High amounts of cholesterol and small amounts of phospholipids and acid mucopolysaccharides are also identified in these sacciform granulated cells.
Phospholipids, mucoproteins and elastin could well increase the viscosity of mucopolysaccharides retaining the slimy secretions on the surface for relatively longer periods. The retention of mucopolysaccharides and elastin on the surface for longer periods that can bind large amounts of water has been correlated to keep the skin moist, facilitating the cutaneous respiration and preventing the fish from desiccation when it is on land. The role of elastin in protecting the fish from various chemical attacks and mechanical stresses is also discussed.
Tryptophan secreted by sacciform granulated cells may have the immunological role facilitating the antibody transfer across the epidermis.     

Carbohydrates in the epidermal mucous cells of a fresh-water fish Mastacembelus pancalus (mastacembelidae,Pisces) as studied by electron-microscopic cytochemical methods

Carbohydrates in the mucous cells of the epidermis of the fish Mastacembelus pancalus were studied by means of electron-microscopic cytochemical methods using physical development procedures. Three types of mucous cells (types A-C) were differentiated on the basis of the reactivities of the secretory products elaborated by them. The carbohydrate contents of mucous globules predominantly comprised sulfate esters and traces of oxidizable vicinal diols in type-A cells, oxidizable vicinal diols in type-B cells, and moderate amounts of both sulfate esters and oxidizable vicinal diols in type-C cells. Glycogen particles were also found to occur in the cytoplasm of these cells, and glycoproteins containing oxidizable vicinal diols were visualized in Golgi cisternae, rough endoplasmic reticulum, nuclear envelopes, and plasma membranes. In the type-A and type-B cells situated in the superficial layers of the epidermis, extensive cisternae of the Golgi apparatus and copious rough endoplasmic reticulum suggested the active syntheses of secretory contents, in contrast to the type-C mucous cells, which displayed poor development of these organelles, in the deeper layers.