Morphological investigation of the gills of the dusky grouper Epinephelus marginatus (Lowe 1834) using gross anatomy and scanning electron microscopy

The current study has designed to investigate the morphological characteristics of the gills of the dusky grouper (Epinephelus marginatus) by gross morphology, scanning electron microscopy, and morphometric analysis. The study was carried out on 10 fresh fish. The fish has four-gill arches on each side. The lengths of the first-, second-, third-, and fourth-gill arches were 5.27, 4.2, 3.2, and 2.8 cm. The gill arches carried a longitudinal band, the bases of the gill filaments, and gill rakers that varied from rectangular to circular shapes. Each gill arch had two main lateral and medial rows and two accessory rows of gill rakers in an alternative manner with each other. The dusky grouper fish had long rakers, whereas the longest one was the lateral rakers of the first arch, which were 467 and 1271.9 μm in width and length. Three types of spines appeared on the gill arches and rakers. The long spine had detected on the apex of the short rakers. Each gill arch carried on its ventral side two rows of gill filaments. The long filaments were at the middle of the gill arch, while the short ones were at the rostral end of the gill arch. The study has demonstrated the taste buds, mucous, chloride cells, and significant features of the epithelial covering of the gill arches and rakers. The morphology of the gills of the dusky grouper indicates the adaptation to their marine environment.     

Surface ultrastructure of gills in relation to the feeding ecology of an angler catfish Chaca chaca (Siluriformes,Chacidae)

Surface ultrastructure of the gills of the angler catfish Chaca chaca was investigated to unravel the adaptive modifications associated with the feeding ecology of the fish. The fish is often found in mud or in soft substrates where they remain buried both for protection and to feed. Gill rakers present on the gill arch in most fish species are absent in this fish. The absence of gill rakers are associated with the feeding habit of the fish and is considered to facilitate the swallowing of captured prey smoothly without any hindrance. Highly corrugated surface of the gill arch and gill filaments could be associated to retain water/mucus to prevent dessicassion of the fish. Papillae like epithelial protuberances each bearing a taste bud at its summit toward the pharyngeal side of the gill arch is associated with the sorting of the food. Large number of mucous goblet cells on the gill arch epithelium are considered to secret copious mucus to lubricate the prey for easy swallowing. In C. chaca the gill septa between gill filaments are reduced. This could enhance the flexibility and permit the free movement of the gill filaments. Extensive secondary lamellae and infrequent mucous goblet cells on secondary lamellae are associated to increase the surface area to enhance efficiency of gaseous exchange.     

Gross morphological and surface ultrastructural investigation on the gills of the European barracuda Sphyraena sphyraena

The present work examined 10 gill systems of European barracuda grossly and by scanning electron microscopy (SEM). Grossly, there were four pairs of the gill arches. The convex border of the gill arch carried the gills filaments, but there were abundance of spines near to its concave border and the gill rakers were absent. Laterally near to the convex border of the gill arch, SEM observations revealed that the first gill arch carried small elliptical, oval, cuboidal, and triangular groups of two shapes of spines; spearhead-like spines and canine-like spines, but the other three gill arches carried larger groups of the same shaped spines with the appearance of waterfalls. Medially near to the convex border of the first gill arch, the canine-like spines were observed only in the form of vertical rectangular groups that adhered in some areas. Laterally near to the concave border, the two shapes of spines were present in the form of longitudinal groups separated by spaces at the first gill arch, but these spaces were absent in the other three gill arches. Medially near to the concave border of the first gill arch, the two shapes of spines were presented in oval groups, while the other three gill arches were covered entirely by cuboidal groups of the two shapes of spines. The absence of the gill rakers in conjunction with an abundance of spines helps the European barracuda to control the food particles from escaping to the gill filaments to prevent its suffocation.     

Surface ultrastructure of the gill filaments and the secondary lamellae of the catfish, Rita rita, and the carp, Cirrhinus mrigala

Surface ultrastructures of gill filaments and secondary lamellae of Rita rita and Cirrhinus mrigala, inhabiting different ecological habitat, were investigated to unravel adaptive modifications. R. rita is a sluggish, bottom dwelling carnivorous catfish, which inhabits regions of river with accumulations of dirty water. It retains its viability for long time if taken out of water. C. mrigala is an active bottom dwelling Indian major carp, which lives in relatively clean water and dies shortly after taken out of water. In R. rita, gill septa between gill filaments are reduced. Microridges on epithelial cells covering gill filaments are often continuous and arranged concentrically. Secondary lamellae are extensive. The epithelium appears corrugated, show irregular elevations and shallow depressions, and microridges on epithelial cells appear fragmented. In C. mrigala, in contrast, the gill septa are extensive. Microridges on epithelial cells covering gill filaments are fragmented. Secondary lamellae are less extensive. The epithelium appears smooth and microridges on epithelial cells are relatively inconspicuous. These differences have been considered adaptive modification in relation to habit and ecological niches inhabited by two fish species. Presence of mucous goblet cells on gill filaments is discussed in relation to their functions including precipitation of the sediments and preventing clogging of gill filaments. Infrequent mucous goblet cells in the epithelium of secondary lamellae in two fish species are considered an adaptation, minimizing thickness of the epithelium to reduce barrier between blood and water for favoring gasses exchange with increased efficiency.     

Biological aspects of the nasal turbinates of the Anatolian shepherd dog captured from Egypt: Using computed tomography,histological, and scanning electron microscopic observations

The current study was designed to describe the nasal turbinates of 15 heads of Anatolian shepherd dogs using the histology and scanning electron microscope. The caudal part of the nasal cavity is almost occupied by the ethmoidal concha that is related to the high dog's smelling. Keratinized stratified squamous epithelial lining of the rostral part of dorsal and ventral concha were interdigitated with the underlying lamina propria, with numerous sebaceous and sweat glands. The pseudostratified squamous epithelium lining of the middle part of the dorsal and ventral conchae had simple seromucous glands. The caudal third of dorsal, ventral, and ethmoidal conchae covered by olfactory epithelium that had three cell types; basal, supporting, and bipolar cells with mucous glands. SEM of the vestibular region shows that the dorsal conchae had a wrinkled surface with microvilli, little olfactory buds, and small sebaceous and sweat glands openings, while the ventral conchae had a lot of filiform-like microvilli. SEM of the respiratory region shows that the dorsal conchae had a little number of seromucous glands and a rosette-shape cilia, while the ventral conchae had numerous cellular cilia that cover all surface. SEM of the fundus region shows that the dorsal conchae had numerous microvilli of ciliated olfactory cells, while the ventral conchae had numerous long microvilli of ciliated olfactory cells. SEM of the ethmoidal nasal conchae shows a dense network of long microvilli of ciliated olfactory cells. We concluded that the morphological features of the dog's nasal turbinates were correlated with their environmental condition.     

Quantitative analysis of keratin filament networks in scanning electron microscopy images of cancer cells

The keratin filament network is an important part of the cytoskeleton. It is involved in the regulation of shape and viscoelasticity of epithelial cells. The morphology of keratin networks depends on post-translational modifications of keratin monomers. In-vitro studies indicated that network characteristics, such as filament crosslink density, determines the biophysical properties of the filament network. This report presents a quantitative method for the morphological analysis of keratin filament networks. Visualization of filaments was based on prefixation extraction of epithelial cells and scanning electron microscopy (SEM). SEM images were processed by a skeletonization algorithm to obtain a graph structure that represents individual filaments as well as their connections. This method was applied to investigate the effects of transforming growth factor α (TGFα) on the morphology of keratin networks in pancreatic cancer cells. TGFα contributes to pancreatic cancer progression and activates signalling pathways phosphorylating keratin monomers. Using this new method, a significant alteration to the keratin network morphology could be detected in response to TGFα.     

Ultrastructure of the ovariole sheath in <i>Diatraea saccharalis</i> (Lepidoptera: Pyralidae)

The ultrastructure of the ovariole sheath along the Diatraea saccharalis ovariole was studied by scanning and transmission electron microscopy. Each ovariole is surrounded by an epithelial sheath, a tunica propria and scattered lumen cells. These three components of the ovariole sheath show different ultrastructural features along the ovariole, in the germarium or in the vitellarium; these differences are more evident in the epithelial sheath cells. The epithelial sheath is composed by two layers of cells, the external one running longitudinally and the internal one running circularly in the ovariole. These cells, in vitellarium, present cytoplasmic bundles of myofilaments that are arranged parallel to the long axis of the cells; these myofilaments are apparently related to the contraction movements of the follicles within the ovariole. The acellular tunica propria, composed of finely filamentous material, is attached to the adjacent follicle cells by adhesive dense plates. Between the epithelial sheath and the tunica propria there is a population of lumen cells, with morphological features of secretory activity     

Light and transmission electron microscopic structure of skin glands and dermal scales of a caecilian amphibian Gegeneophis ramaswamii,with a note on antimicrobial property of skin gland secretion

Amphibian skin secretions contain a variety of bioactive compounds that are involved in diverse roles such as communication, homeostasis, defence against predators, pathogens, and so on. Especially, the caecilian amphibians possess numerous cutaneous glands that produce the secretory material, which facilitate survival in their harsh subterranean environment. Inspite of the fact that India has a fairly abundant distribution of caecilian amphibians, there has hardly been any study on their skin and its secretion. Herein, we describe, using light microscopy and electron microscopy, two types of dermal glands, mucous and granular, in Gegeneophis ramaswamii. The mucous glands are filled with mucous materials. The mucous‐producing cells are located near the periphery. The granular glands are surrounded by myoepithelial cells. A large number of granules of different sizes are present in the lumen of the granular gland. The granule‐producing cells are present near the myoepithelial lining of the gland. There are small flat disk‐like dermal scales in pockets in the transverse ridges of the posterior region of the body. Each pocket contains 1–4 scales of various sizes. Scanning electron microscopic (SEM) study of the skin surface showed numerous funnel‐shaped glandular openings. The antibacterial activity of the skin secretions was revealed in the test against Escherichia coli, Klebsiella pneumoniae, and Aeromonas hydrophila, all gram‐negative bacteria. SEM analyses confirm the membrane damage in bacterial cells on exposure to skin secretions of G. ramaswamii.     

In vitro confocal imaging of the rabbit cornea

We were able to observe in vitro the fine structure of the rabbit cornea using a laser scanning confocal microscope, especially in the regions between Descemet's membrane and the epithelial basal lamina. We observed submicrometre filaments throughout the stroma with high concentrations adjacent to Descemet's membrane, and found extensive interconnecting processes between stromal keratocytes. There are numerous regions containing nerve plexuses in the stroma. We found a deeply convoluted basal lamina adjacent to the epithelium, and observed regions containing junctions between endothelial cells in fluorescent images of rabbit corneas stained with the actin-specific compound fluorescein phalloidin.     

Scanning electron microscopy and morphometric analysis of superficial corneal epithelial cells in dromedary camel (Camelus dromedarius)

It is likely that superficial corneal epithelial cells (SCECs) of the dromedary camels have a significant role in their survival at arid and semiarid regions. To the best of our knowledge, SCECs of camels' eyes have not been characterized previously using scanning electron microscopy (SEM), combined with morphometric analysis. Therefore, in the current study, we aim to describe the shape, topographical distribution, and density of SCECs associated with morphometric analysis using SEM. Twelve healthy adult camels' corneas were obtained immediately after slaughter. Each cornea has been divided into nine parts: central (C), middle dorsal (MD), middle ventral (MV), middle nasal (MN), middle temporal (MT), peripheral dorsal (PD), peripheral ventral (PV), peripheral nasal (PN), and peripheral temporal (PT). SCECs were distinguished and characterized into light, medium, and dark mosaics. The polygonal cells have been externally covered with microplicae that were more numerous above the light cells. The topographic distribution of light, medium, and dark cells revealed a well-defined concentration of light cells in excess of other cells in all parts as follows: PV (92.5%), PN (78.5%), MN (78%), MT (74.7%), PD (73.8%), PT (70.7%), MV (68.7%), MD (66.3%), and C (19.3%). The PV part recorded the highest density of light cells, while the C portion showed the lowest density for the same cells. We concluded that the light cells extensively predominate in all parts of the camels' cornea except the C part, indicating an adaptive modification to the harsh environment. Additionally, the PV and PN parts represent the permanent and endogenous source as well as a proliferative reserve for SCECs in dromedary camel.     

Fine structural aspects of secretion and extrinsic innervation in the olfactory mucosa.

The mucus at the surface of the olfactory mucosa constitutes the milieu in which perireceptor events associated with olfactory transduction occur. In this review, the ultrastructure of olfactory mucus and of the secretory cells that synthesize and secrete olfactory mucus in the vertebrate olfactory mucosa is described. Bowman's glands are present in the olfactory mucosa of all vertebrates except fish. They consist of acini, which may contain mucous or serous cells or both, and ducts that traverse the olfactory epithelium to deliver secretions to the epithelial surface. Sustentacular cells are present in the olfactory epithelium of all vertebrates. In fish, amphibia, reptiles, and birds, they are secretory; in mammals, they generally are considered to be "non-secretory," although they may participate in the regulation of the mucous composition through micropinocytotic secretion and uptake. Goblet cells occur in the olfactory epithelium of fish and secrete a mucous product. Secretion from Bowman's glands and vasomotor activity in the olfactory mucosa are regulated by neural elements extrinsic to the primary olfactory neurons. Nerve fibers described in early anatomical studies and characterized by immunohistochemical studies contain a variety of neuroactive peptides and have several targets within the olfactory mucosa. Ultrastructural studies of nerve terminals in the olfactory mucosa have demonstrated the presence of adrenergic, cholinergic and peptidergic input to glands, blood vessels, and melanocytes in the lamina propria and of peptidergic terminals in the olfactory epithelium. The neural origins of the extrinsic nerve fibers and terminals are the trigeminal, terminal, and autonomic systems.     

Ultrastructure of Tracheal Surface Liquid: Low-Temperature Scanning Electron Microscopy

A layer of liquid lines the airways in the lung. Previous microscopic studies have suggested that it is in two phases, with a mucous gel lying above a periciliary sol. However, shrinkage artifacts due to chemical fixation, dehydration, and drying have prevented reliable estimates of the depth of these layers. To avoid such problems, we have studied the surface liquid of bovine trachea by low-temperature scanning electron microscopy (LTSEM). A polished copper probe cooled to liquid nitrogen temperature was applied to the mucosal surface of sheets of excised tracheal epithelium to effect rapid freezing of surface liquid. Tissue sheets were then mounted in an LTSEM (AMRay 1000A with Biochamber) which maintains samples at -180°C with a Joule-Thompson refrigerator built into the stage. Tissues were fractured at right angles to the epithelial surface, coated with gold, and viewed, all at 10^?5 to 10^?6 torr without transfer through air. The sample was stable under the electron beam at accelerating voltages up to 20 kV. Epithelial features (nuclei, cilia, microvilli, mucous granules) were well preserved. The mucosal surface of the cells was covered with material on the order of 8 μm in depth. The mucous gel and periciliary sol could be seen as distinct layers and could be distinguished by the size and pattern of ice crystal voids generated by radiant-etching of the fractured surface of the sample.     

Unravelling the structure of the lamellipodium

Summary Pushing at the cell front is the business of lamellipodia and understanding how lamellipodia function requires knowledge of their structural organization. Analysis of extracted, critical-point-dried cells by electron microscopy has led to a current dogma that the lamellipodium pushes as a branched array of actin filaments, with a branching angle of 70 degrees , defined by the Arp2/3 complex. Comparison of different preparative methods indicates that the critical-point-drying-replica technique introduces distortions into actin networks, such that crossing filaments may appear branched. After negative staining and from preliminary studies by cryo-electron tomography, no clear evidence could be found for actin filament branching in lamellipodia. From recent observations of a sub-class of actin speckles in lamellipodia that exhibit a dynamic behaviour similar to speckles in the lamella region behind, it has been proposed that the lamellipodium surfs on top of the lamella. Negative stain electron microscopy and cryo-electron microscopy of fixed cells, which reveal the entire complement of filaments in lamellipodia show, however, that there is no separate, second array of filaments beneath the lamellipodium network. From present data, we conclude that the lamellipodium is a distinct protrusive entity composed of a network of primarily unbranched actin filaments. Cryo-electron tomography of snap-frozen intact cells will be required to finally clarify the three-dimensional arrangement of actin filaments in lamellipodia in vivo.     

Application of stereologic methods to stratified gingival epithelia

Stereologic point-counting procedures were applied to estimate quantitative tissue and cytoplasm parameters of the oral and the junctional epithelium of the human gingiva. Three gingival biopsies of female children, obtained and processed under standardized conditions, served (1) to determine the minimal sample size of cytoplasmic area required for estimating representative volumetric parameters in different strata of both epithelia, and (2) to compare average volume and surface density data derived from standardized samples in epithelial cross sections cut at two planes orientated perpendicular to each other. Sampling of electron micrographs, performed at two levels of magnifications according to a systematic stratified random sampling procedure consisted of recording field strips parallel to the epithelial or tooth surface within each of the various strata. The optimal sample size required varied for different organelles and epithelial strata. Minimal sample size of cytoplasmic area per biopsy were 300–440 μm² for basal, and 450 μm² for stratum spinosum cells of oral epithelium, and 130–185 μm² for basal cells of the junctional epithelium. Average morphometric parameters for gross tissue components (cells, nuclei, cytoplasm, intercellular space) which resulted from analysing 4900 μm² tissue area in each of the epithelia in each biopsy, were almost identical when determined in two different section planes. Volume and surface density data of cytoplasmic organelles (mitochondria, rough and smooth endoplasmic reticulum, cytoplasmic filaments) resulting from a sample of 280 μm² cytoplasmic area per stratum and biopsy, revealed differences of varying magnitude, when determined in the two section planes. These differences were markedly smaller than those between comparable strata of both epithelia. Discussion of sampling procedures concluded that the type of sampling used for the present study is suitable for comparative morphometric studies.     

Brush-like fibers on the human chromosome periphery

A brush-like border apparently composed of fibers protruding from metaphase chromosomes of human lymphocytes was observed for the first time using transmission (TEM) and scanning electron microscopy (SEM). On the basis of size and sensitivity to colcemid, the fibers may be related to microtubules and spindle organization. The brush-like fibers were observed when chemically fixed metaphase chromosome spreads were placed on glass slides and maintained under "wet" conditions (not allowed to air dry after fixation for conventional cytogenetic protocols) until postfixation protocols for TEM and SEM were performed. The purpose of this study was to establish the occurrence of the brush-like fibers and to determine the effects of colcemid on these fibers.     

Healing of cutaneous wounds in a freshwater teleost, Labeo rohita: scanning electron microscopical investigation

In this study, healing of cutaneous wounds in Labeo rohita using scanning electron microscope is reported. Wound area could be divided into three regions. Immediately after infliction of wound, edges retract exposing underlying tissues in wound gap (Region I). Simultaneously, at region close to wound edge (Region II), mucous goblet cell openings are observed with copious mucous secretions. Within 1 h, Region I gets covered by mucous secretions, and epidermis at edges starts migrating. Opposing fronts gradually advance and by 4-6 h come in contact to epithelialize wound gap. Zone of contact of fronts is demarcated by epidermal ridge, which is relatively prominent at 8 h. It gradually diminishes and is not distinguished at 24 h and afterward. At 1-4 h, microridges on epithelial cell surfaces appear irregularly arranged, widely spaced, short with abrupt ends at Region I; relatively extensive at Region II; and similar to those in controls at region surrounding Region II (Region III). At 12 h and afterward, microridges appear similar to those in controls at Regions I and II. At 1-2 h, isolated swollen epithelial cells, often in process of detachment and exfoliation at surface, are observed at Regions I and II. Such cells are infrequent at 8 h and afterward. Region I covered by migrated epidermis appears trough like at 4 h to 2 days, level of which gradually rises and at Day 4, surface of epidermis appears at a level similar to that at Regions II and III. Changes have been associated with the imbalance of osmotic homeostasis due to disruption of barrier between internal and external environment of skin.     

Morphological organization of the dorsal protuberance of Linepithema humile (Mayr, 1868) ant's larvae (Hymenoptera, Formicidae)

The Argentine ant Linepithema humile is an important invasive species because of the levels of infestation that it can reach; however, there is little information about its presence, histological organization, and function of the dorsal protuberance, which is found exclusively in their larvae. The objective of this study was to describe it in L. humile through scanning electron microscopy and transmission electron microscopy, bringing information about this structure. The epidermis of these larvae have cuticles covering the whole body, and is formed by a sequence of overlapping lamellas where the inner ones were thicker and presented lower electron density, whereas the outer ones were thinner and highly electron dense. Pores or pore-like channels were not observed. A thick and acellular region composed of granular material was found under the cuticular layer. Out of this region, the flattened epidermic cells formed an epithelial layer. For the dorsal protuberance region, these cells become prismatic, and similarly to the cuticle, presents significant thickening. These cells presented extended microvilli, as well as a great amount of lamellar rough endoplasmic reticulum. Under this epithelium was observed a concentration of fat body cells, more numerous in the dorsal protuberance region. This study indicated that the dorsal protuberance present in the first segment of L. humile larvae has apparently no secretory function because no pores were found. This fact allowed to conclude that in L. humile larvae the dorsal protuberance would have the function to make it easier for the worker ants to carry them within the colony.     

Light and transmission electron microscopy study of the peripheral olfactory organ of the guppy, Poecilia reticulata (Teleostei, Poecilidae)

A study of the peripheral olfactory organ, with special attention to the olfactory epithelium, has been carried out in the guppy (Poecilia reticulata). Guppy is well known to have a vision-based sexual behavior. The olfactory chamber caudally opens directly in an accessory nasal sac, which is bent medially and gives rise to two recesses that can be considered secondary accessory nasal sacs, antero-medial and postero-medial, respectively. The sensory epithelium, which lines only the medial wall of the nasal cavity, is basically flat rising in a very low lamella only in the posterior part. The olfactory receptors are not evenly distributed in the olfactory mucosa, but aggregate in shallow folds separated by epithelial cells with evident microridges. Ciliated olfactory sensory neurons and microvillous olfactory sensory neurons are clearly identified by transmission electron microscopy (TEM). Scarce crypt olfactory neurons are found throughout the sensory folds. The nasal sacs indicates the capacity to regulate the flow of odorant molecules over the sensory epithelium, possibly through a pump-like mechanism associated with gill ventilation. The organization of the olfactory organ in guppy is simple and reminds what is found in early posthatching stages of fish which at the adult state have a well developed olfactory organ. This simple organization supports the idea that the guppy rely on olfaction less than other fish species provided with more extended olfactory receptorial surface.     

A balanced technique for preparation of specimens from pathogenicity studies for scanning electron microscopy

This paper reports our experiences with preparing delicate biological specimens for scanning electron microscopy. Three different washing methods were evaluated: One method allowed the analysis of the location of the bacterium Mycoplasma mobile on piscine gill epithelium and the optimal evaluation of histopathologic changes caused by this microbe. These results were achieved when specimens were washed three times in a cacodylic acid buffer after completion of the in vitro infection experiment in gill explant cultures. We also found that of three different concentrations of glutaraldehyde, a fixation with a 1.5% solution was sufficient to achieve excellent structural preservation, even without using post fixation in osmium tetroxide. Furthermore, this study showed that the use of acetone-carbon dioxide in the critical point drying procedure resulted in well-preserved piscine gill epithelium and mycoplasmas. Finally, long-term storage of tissue specimens in 0.1 M cacodylic acid buffer is possible if the buffer is changed on a monthly basis to avoid growth of unwanted microorganisms, such as fungi.