Histochemistry and the structure of the epidermis of a fresh-water teleost Noemacheilus botia (Hamilton-Buchanon) (Cobitidae, Pisces)

Functional organization and the histochemical nature of the various cellular components of the epidermis of Noemacheilus botia are described. The various histochemical techniques reveal the basic proteinaccous nature of the outer free margins of the polygonal cells of the most superficial layer of the epidermis. These cells remain metabolically active as revealed by their healthy nuclei and are not sloughed off at the surface. the lateral cell membranes of these cells are fused together forming a continuous barrier which plays important role in water proofing the skin. In addition the polygonal cells in the most superficial layer also undergo the process of mucogenesis synthesizing sulphated acid mucopolysaccharides which may ultimately form a part of the contents of the protective extracellular cuticular coat.     

Protein and carbohydrate histochemistry in relation to the keratinization in the epidermis of Barbus sophor (Cyprinidae, Pisces)

The distribution of protein and carbohydrate constituents in the epidermis of Barbus sophor is described in order to give a better understanding of its cellular organization and physiology.
Various cytochemical techniques show the keratinized nature of the outer free margins of the polygonal cells in the most-superficial layer. These contain appreciable amounts of cysteine bound sulphydryl groups, basic proteins, protein bound NH₂ groups, ribonucleic acid and calcium and give a strong Papanicolaou's reaction. Absence of cystine bound disulphide groups suggests that the cornified layer in B. sophor is probably mechanically weak as adjacent keratin chains remain unbonded. The polygonal cells showing keratinization at the outer free margins remain metabolically active and are not sloughed off at the surface. This is in contrast to the keratinized epidermis of other teleosts so far reported in which the keratinized cells are dead and are sloughed off at the surface.
In addition to keratinization the polygonal cells undergo mucogenesis synthesizing sulphated acid mucopolysaccharides.
The presence of eosinophilic granular cells in the epidermis is interesting. The possible role of these cells in the protection of the epidermis has been discussed. The epidermis on the inner surface of the scale is very thin so it may not have much protective significance in these areas.     

Structure of the integument of a fresh-water teleost,Bagarius bagarius (Ham.) (Sisoridae,pisces)

The skin of Bagarius bagarius (Ham.) is devoid of scales but is rough due to the presence of numerous pentagonal epidermal elevations, which are separated by deep furrows at regular intervals. These elevated pentagonal regions of the epidermis are covered by dead cornified cells in the form of caps. As the old cap goes off a new one is formed by the death of the underlying epidermal cells. The middle layer of the epidermis is mainly composed of well defined polygonal cells. Their cytoplasm is granular in nature and give reactions for protein bound sulphydryl groups. The stratum germinativum is composed of two types of basal cells, the columnar cells and the spherical cells. The flask shaped mucous glands are restricted to the epidermal furrows and secrete either neutral or acidic mucopolysaccharides. Certain large specialysed granular cells are found in the epidermis which are grouped around the taste buds. These specialysed cells may be the photocytes. Two layers of the dermis can be distinguished—the relatively thin stratum laxum and the thick stratum compactum. Dermal papillae mainly support the taste buds. The pigment cells are arranged in two layers in the dermis. The subcutis is composed of loose connective tissues, richly infiltrated with the fat cells, nerves and blood capillaries.     

Enzymes in the epidermis of Natrix piscator during its sloughing cycle

Acid phosphatase, non-specific esterase, alkaline phosphatase, monoamine oxidase and true lipase activities, in the epidermis of Natrix piscator in different stages of the sloughing cycle, have been localized using various histochemical techniques.
Different layers in scale epidermis have staining properties similar to corresponding layers in hinge epidermis.
Acid phosphatase and non-specific esterase activity in cell layers undergoing keratinization, and the lacunar tissue undergoing disintegration are associated with hydrolytic and catabolic wasting processes involving cell death. The activity of these enzymes in the clear layer is associated with the breaking down of the cementing substance resulting in the separation of clear layer from underlying tissue and facilitating the shedding of old slough.
Alkaline phosphatase activity in the stratum germinativum and undifferentiated epidermal cells has been associated with cell proliferation and differentiation. The presence of alkaline phosphatase in the lacunar tissue and clear layer has been correlated with the synthesis of mucopolysaccharides in these layers.
Monoamine oxidase and true lipase activity could not be located in the epidermis at any stage of the sloughing cycle.     

Lipids in the skin of a cat-fish Heteropneustes fossilis (Bloch) (Heteropneustidae,Pisces)

Summary Localization and characterization of different lipids in the cellular constituents of the skin of Heteropneustes fossilis has been made using several histochemical techniques.High contents of cholesterol, its esters and phospholipids have been correlated with the metabolically active state of the basal cells undergoing cell proliferation and differentiation.The polygonal cells in the outermost layer of the epidermis, though rich in phospholipid contain small amounts of cholesterol and its esters. This has been correlated with the metabolically less active state of these cells.Neutral lipids and phospholipids in the polygonal cells of the outermost layer may contribute to the contents of surface slime and act as an efficient barrier for the penetration of water through the skin.The deposits of neutral lipids in the subcutis may provide energy during the period of fasting, act as a barrier for water diffusion through the skin and serve as shock absorbing pads protecting the fish from mechanical injury.Supported by a post doctoral fellowship sponsored by the Council of Scientific and Industrial Research, Government of IndiaSupported by a research scholarship sponsored by Banaras Hindu University     

Carbohydrate histochemistry of the epidermis of Natrix piscator during its sloughing cycle

Carbohydrate histochemistry of the scale and hinge epidermis of the chequered water snake, Nalrix piscator , throughout the sloughing cycle, has been described. The small amount of mucopolysaccharide present in the Oberhautchen, mesos layer, α-layer and β-Mayer (in its initial stage of differentiation) is comparable with that in amphibian epidermis and the epidermis of certain freshwater fish undergoing keratinization. Moderate amounts of mucopolysaccharide in the lacunar tissue and clear layer may protect against environmental pathogens and retain water to protect the epidermis from desiccation. Mucous cells could not be located in the epidermis throughout the sloughing cycle, contrary to some previous observations that they occur in the hinge region. The general absence of glycogen in the epidermis in most stages of the sloughing cycle suggests that the glycogen metabolized in the epidermis is utilized immediately, in view of the high energy requirements of proliferation and differentiation. Accumulation of glycogen granules in the presumptive α-layer in stage 2 and in the clear layer, presumptive Oberhautchen and presumptive β-Mayer in stage 3 is correlated with low energy requirements, indicating a slowing down of the process of keratinization of cells in these layers.     

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.     

The spinal nerves innervate putative chemosensory cells in the ventral skin of desert toads, Bufo alvarius

Toads normally obtain water by absorption across their skin from osmotically dilute sources. When hyperosmotic salt solutions are presented as a hydration source to dehydrated desert toads, they place the ventral skin onto the source but soon afterwards escape to avoid dehydration. The escape behavior coincides with neural excitation of the spinal nerves that innervate putative chemosensory cells in the ventral skin. In the present study, fluorescent dye translocated through the spinal nerves to those receptor cells in the epidermis was photoconverted in the presence of 3, 3'-diaminobenzidine tetrahydrochloride for electron-microscopic observation of the cells and associated nerve terminals. Most of the photoconverted cells were located in the deepest layer of the epidermis, with some being in more intermediate layers. No labeled cell was seen in the outermost layer of living cells. In desert toads, flask cells and Merkel cells are occasionally seen in the epidermis. An association of nerve fibers with these epidermal cells has been reported in some species of the anurans. In the present study, however, the cytological features of the photoconverted cells are neither reminiscent of flask cells nor Merkel cells, but are similar to those of surrounding epithelial cells in each layer of the epidermis. We hypothesize a sensory function for these cells, because they have a close association with nerve fibers and participate in the transepithelial transport of salts that must pass through all cell layers of the skin.     

Structure of the skin of an air-breathing mudskipper, Periophthalmus magnuspinnatus

The epidermis of the mudskipper Periophthalmus magnuspinnatus consisted of three layers: the outermost layer, middle layer and stratum germinativum. Extensive vascular capillary networks were present near the superficial layer of epidermis and outermost layer. The diffusion distance between the vascular capillaries and the surface of epidermis was c . 1.5 ± 0.9μm. The middle layer consisted of small or voluminous cells swollen by epidermal cells. Due to the swollen cells, the thickness of the epidermis increased and the epidermis appeared web-like. The swollen cells contained tonofilaments, lucent contents and desmosomes. Fine blood capillaries were also discernible in this layer. Well-developed lymphatic spaces containing lymphocytes existed in the stratum germinativum. Numerous blood capillaries were present under the basement membrane. The dermis consisted of a stratum laxum and stratum compactum, and there was a definite area with acid mucopolysaccharides and a small scale in the stratum laxum. The skin had an epidermal pigment cell, dendritic melanophores (-cytes) containing melanin granules within their cytoplasm, and two kinds of dermal pigment cells, melanophores and colourless pigments containing reflecting platelets.     

Glycogen Distribution in Relation to Epidermal Cell Differentiation During Embryonic Scale Morphogenesis in the Lizard Anolis lineatopus

The differentiation of the epidermis during scale morphogenesis in the lizard Anolis lineatopus has been studied by autoradiographic and immunocytochemical techniques and by electron microscopy, in relation to mitotic activity and to the distribution of glycogen. The flat embryonic epidermis of the early embryo is transformed into symmetric epidermal papillae which progressively become asymmetric and eventually form scales with stratified epidermal and peridermal layers. Papilla asymmetrization and epidermal stratification derive from cell hypertrophy and multiplication in the “basal hypertrophic layer of the forming outer side of scales” (BLOS). Glycogen is scarce or absent during early stages of epidermis development. In the dermis no glycogen is found at any stage of scale morphogenesis. Glycogen particles 25–40 nm in size accumulate in hypertrophic basal cells and peridermal cells during scale development. Conversely cells in the forming inner side of scales do not accumulate glycogen, divide less frequently than in the outer side and do not form a β–keratinized layer. It is suggested that an osmotic effect related to glycogen deposition causes increased hydration of the BLOS, whose cells become swollen and contribute to the asymmetrization of the epidermal papillae. Glycogen decreases in suprabasal differentiating cells and disappears from the BLOS at the stage of complete keratinization of the scale, around the period of hatching. Terminal differentiation in the peridermis and suprabasal epidermal layers takes place by cell flattening and condensation of the nucleus and cytoplasm as typical for apoptotic cells.     

The conversion of injected glucose into renal glycogen and mucopolysaccharides

Summary Tritiated glucose has been injected into rabbits in various states of hydration. The renal papilla of all animals showed an uptake of the label, converted into glycogen, and into mucopolysaccharides, in a manner dependent on the water balance of the animal.In papillae of control animals, the glycogen of the collecting duct epithelial cells and the mucopolysaccharides of the interstitium were labelled.In papillae of animals in an aqueous diuresis, the collecting duct glycogen was lightly labelled and there was no label over the interstitium.Antidiuretic hormone caused a diversion of label from the collecting ducts into interstitial mucopolysaccharides.The significance of these findings, with respect to renal concentrating ability, is discussed.The author wishes to thank Dr. M. K. S. Hathorn for help with the statistical analysis. Mr. K. Gamblin and Mr. P. L. Hyam gave valuable technical assistance. This research formed a part of the work approved for the degree of Ph. D. (London).     

On the presence of high glycogen concentration and intermediate filaments in the flat cells of the electroreceptive epidermis of mormyrid fish

The cytoplasm of the flat cells of the electroreceptive epidermis of Mormyrids was examined in the light and the electron microscope, in order to reveal the presence of glycogen and to study its distribution. In the electroreceptive epidermis, which consists of three layers, the periodic acid Schiff reaction used to stain polysaccharides is strongly positive in the superficial polyhedral cells and in the flat cells of the intermediate layer. Polysaccharides are absent in the basal polyhedral cells. Pre-incubation with alpha-amylase shows that glycogen is present only in the intermediate cell layer. In the electron microscope, after reaction with periodic acid, thiocarbohydrazide and silver proteinate, glycogen is seen in the form of rosettes of monoparticles. These rosettes occupy both the central region of the cytoplasm of these cells, and the more peripheral parts, where alignments of desmosomes are found. In the cytoplasm of certain flat cells, the rosettes are grouped to form accumulations of glycogen which cover several mu2. Observation in the electron microscope reveals that in addition to glycogen, these cells contain tonofilaments or intermediate filaments, common to epithelial cells, which may group themselves in bundles. Glycogen and the intermediate filaments are thus the principal constituents of the cytoplasm of the flat cells of the electroreceptive epidermis of Mormyrids. The possible role of the filaments, and especially of the glycogen which is a polysaccharide high in energy, in the flat cells which apparently have a low metabolic rate, is discussed.     

Differentiation of Wing Epidermal Scale Cells in a Butterfly Under the Lateral Inhibition Model - Appearance of Large Cells in a Polygonal Pattern

Cellular pattern formations of some epithelia are believed to be governed by the direct lateral inhibition rule of cell differentiation. That is, initially equivalent cells are all competent to differentiate, but once a cell has differentiated, the cell inhibits its immediate neighbors from following this pathway. Such a differentiation repeats until all non-inhibited cells have differentiated. The cellular polygonal patterns can be characterized by the numbers of undifferentiated cells and differentiated ones. When the differentiated cells become large in size, the polygonal pattern is deformed since more cells are needed to enclose the large cell. An actual example of such a cellular pattern was examined. The pupal wing epidermis of a butterfly Pieris rapae shows a transition of the equivalent-size cell pattern to the pattern involving large cells. The process of the transition was analyzed by using the method of weighted Voronoi tessellation that is useful for treatment of irregularly sized polygons. The analysis supported that the pattern transition of the early stage of the pupal wing epidermis is governed by the lateral inhibition rule. The differentiation takes place in order of largeness, but not smallness, of the apical polygonal area in the differentiating region of the pupal wing.     

中国黄耆属簇毛黄耆亚属的叶表皮特征及其系统学意义

在光学显微镜和扫描电镜下,观察了国产黄耆属簇毛黄耆亚属16种1变种的叶表皮特征。结果表明,气孔器在各种植物的上、下表皮均有分布,多为无规则型,也有不等细胞型;叶表皮细胞形状有不规则形与多边形,表皮细胞垂周壁有平直、浅波状、波状或深波状。叶表皮细胞形状与垂周壁的式样可以分为四种类型,这四种类型与亚属的分组有一定的对应关系。在扫描电镜下可见表皮细胞上有角质层,气孔下陷,气孔的外拱盖及其内缘特征在亚属内都比较一致。表皮细胞角质层的纹饰在个别类群中有一定的变异,对种类的鉴定有一定的意义。     

Functional organization of the skin of the ‘Green-puffer fish’Tetraodon fluviatilis (Ham.-Buch.) (Tetraodontidae,Pisces)

Summary The present study concerns the functional organization of the skin ofTetraodon fluviatilis. The epidermis consists of five different types of cells — the flask-shaped mucous cells, the eosinophilic granular cells, the sacciform granulated cells, the vesicle containing granulated cells, and the polygonal cells. A thin noncellular layer, the cuticle found on the surface of the skin, is probably secreted from the polygonal cells in the outermost layer of the epidermis. A,well-defined lymphatic plexus exists between the cells of the basal layer.Numerous triradiate calcareous spines are embedded within elastic connective tissue pockets in the thick dermis. These pockets are filled with an amorphous, acellular, PAS positive material, and are richly supplied with fine blood capillaries. A histomorphologic basis for the erection of the spines and various structural modifications in the skin facilitating its enormous stretching under inflated conditions of the fish are discussed.Abbreviations Used BCA blood capillary - BM basement membrane - BC basal cell - BL basal layer - CFB collagen fiber bundle - CTB connective tissue band - DER dermis - EGC eosinophilic granular cell - EPD epidermis - FB fibroblasts - FC fat cell - L lymphocyte - LS lymphatic space - MC mucous cell - ML middle layer - MUS muscle - MYS myocommata - NV nerve - OL outermost layer - PCB black pigment cell - PCY yellow pigment cell - PEC polygonal epidermal cell - SCT subcutis - SGC sacciform granulated cell - SP spine - STC stratum compactum - STL stratum laxum - VGC vesicle containing granulated cell - VS vertical strand This investigation was supported by a research grant No. 38(131)/72-GAU-II from the Council of Scientific and Industrial Research and a financial assistance grant for teachers No. F. 6(4626) 72-(SF-1), from the University Grants Commission, Government of India, New Delhi.     

The relationship between nitrogen output and changes in mucous cell function in the amphibious teleost Blennius pholis L. during aerial exposure

The numbers of mucous cells in the epidermis of the head and body and in the surface of the gill arch of the amphibious blenny, Blennius pholis L., were estimated on fish immersed in sea water and after 4 h aerial exposure. During emersion there appeared to be a considerable reduction in the frequency of epithelial mucous cells in the areas studied, although counts for the head epidermis were somewhat variable. A concomitant decline in the number of cells thought to be actively secreting was also recorded in tissue samples from both the head and gills, while in the body epidermis the potential for mucus-secretion was maintained close to the levels observed in immersed fish.
Histochemical studies revealed epidermal mucous cells containing either sialylated acid mucopolysaccharides or neutral mucins, or a mixture of these, in both the head and the body, whereas in the gill arch epithelia there were, in addition, cells containing sulphated acid mucopolysaccharides. After emersion, a disproportionate loss of cells containing neutral and sialylated mucus from the gill epithelia resulted in an increase in the proportion of secreting cells staining positively for sulphated acid mucopolysaccharides.
The results of this study are discussed in relation to nitrogenous excretion during aerial exposure.     

Histochemical studies on the body wall of nematodes: Haemonchus contortus (Rud., 1803) and Xiphinema insigne Loos, 1949.

Histochemical studies on the body wall of Haemonchus contortus (Rud.) and Xiphinema insigne Loos have been made. In H. contortus, the cuticle is mainly proteinous in nature. The lipids and PAS-postive materials are only present in cortical layers. In addition, haemoglobin and acid phosphatase are also present. The hypodermis shows the presence of glycogen, lipids, RNA, acid and alkaline phosphatases. The oval dense body is composed of keratinous and collagenous proteins associated with acid mucopolysaccharides. Muscles carry a greater concentration of glycogen granules and phospholipids. In X. insigne, the cuticle is rich in sudanophilic lipids. The cuticle also consists of weakly acidic mucopolysaccharides. Hypodermis and muscles contain lipids and glycogen. In addition, hypodermis also consists of acidic mucopolysaccharides. The functional significance of these components has been fully discussed.     

Changes in the specific cytochemical properties of rat decidual cells during differentiation

The antimesometrial part of rat's decidua of 8-9 day of gestation was divided into three parts: epithelial, transition and basal zones. Cells of either zone have their own morphological and cytochemical features. Cells of the epithelial zone are characterized by synthesis of tissue specific antigens and fat (fatty acids). Cells of the transition zone synthesize glycogen. Acid mucopolysaccharides are located in the basal zone only. Decidual cells do not synthesize cholesterol. Con A receptors are localized on the surface of cells of the basal and transition zones and disappear from the surface of epithelial zone cells. It is concluded that differentiation of large decidual cells of the antimesometrial part of rat's decidua are accompanied by a significant change in cytochemical features of cell precursors the synthesis of acid mucopolysaccharides and glycogen stops, while tissue specific antigens and fat (fatty acids) start their syntheses, Con A receptor disappear.     

Histochemistry of the glands associated with the reproductive tract ofLymnaea stagnalis

Summary The albumen gland, the muciparous gland and the oöthecal gland of female genital tract of Lymnaea stagnalis, collected in spring, autumn and winter have been studied.The reactions for polysaccharides, proteins and RNA have been performed in order to characterise the secretion of the glands.The albumen gland secretion consists almost exclusively of slightly acid polysaccharides whose histochemical reactions, according to Lison and Grainger and Shillitoe confirm the presence of galactogen. Proteins are also present in the secretion. The muciparous gland secretion consists of strongly acid mucopolysaccharides (non sulphated) produced by large cells among which small cells containing sulphated mucopolysaccharides are present.In the oöthecal gland two zones are present, one with a single type of cells containing acid mucopolysaccharides, and the other with two different types of cells: the first with mucopolysaccharides and the second with sulphated mucopolysaccharides, proteins and glycogen at the basis of the cell. Sialic acids are not present in the secretion of the glands studied.The polysaccharidic composition of the secretion of the glands is different from gland to gland. The secretion of the glands gradually changes and gets acid according to the composition of the various membranes and envelopes wrapping up the eggs.     

Histochemical studies of mucosubstances and lipids in normal human oesophageal epithelium

Synopsis The histochemistry of lipids and mucosubstances in normal human oesophageal epithelium were studies in biopsies obtained from 24 patients undergoing investigations of the upper gastro-intestinal tract. Neutral fat droplets, 1–2 gmm in diameter, were present in all layers, the greatest number being in the functional layer cells. No unsaturated lipids or fatty acids were demonstrable. Much glycogen was present in the cytoplasm of the prickle and functional cell layers as demonstrated by PAS (and diastase digestion) techniques. The intercellular space of the most superfical of the functional cell layer contained neutral and sialic acid-rich acid mucopolysaccharides. These may be important in protecting the epithelium against physical and chemical trauma.