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1.
The existence of a layer of mucus covering the gill lamellae of healthy rainbow trout (Oncorhynchus mykiss) was investigated. Using cryo-scanning electron microscopy, a smooth, undulating, thin layer was observed which completely covered gill filaments and lamellae, thereby obscuring epithelial microridges. After processing cryopreserved gill arches in glutaraldehyde for conventional scanning electron microscopy, the layer was no longer present and epithelial microridges were clearly visible. The identity of this layer was investigated using cryopreserved gills which were treated in one of two ways. First, gills were incubated with a rabbit antiserum to gill mucus, with normal rabbit serum, or with phosphate-buffered saline. Following fixation in glutaraldehyde and processing, only the gill tissue incubated with the mucus-specific antiserum was still covered with the smooth layer. The layer was also retained on the gills of fish anesthetized in a solution containing mucusspecific antiserum and then processes in glutaraldehyde for conventional scanning electron microscopy. The tenacious nature of the mucous layer was demonstrated by its stability following exposure to formalin and a cationic detergent. Second, the presence of this layer was confirmed on gill tissue which was cryopreserved, followed by freeze-substitution and vapor fixation, and then examined by transmission electron microscopy.  相似文献   

2.
The existence of a layer of mucus covering the gill lamellae of healthy rainbow trout (Oncorhynchus mykiss) was investigated. Using cryo-scanning electron microscopy, a smooth, undulating, thin layer was observed which completely covered gill filaments and lamellae, thereby obscuring epithelial microridges. After processing cryopreserved gill arches in glutaraldehyde for conventional scanning electron microscopy, the layer was no longer present and epithelial microridges were clearly visible. The identity of this layer was investigated using cryopreserved gills which were treated in one of two ways. First, gills were incubated with a rabbit antiserum to gill mucus, with normal rabbit serum, or with phosphate-buffered saline. Following fixation in glutaraldehyde and processing, only the gill tissue incubated with the mucus-specific antiserum was still covered with the smooth layer. The layer was also retained on the gills of fish anesthetized in a solution containing mucusspecific antiserum and then processes in glutaraldehyde for conventional scanning electron microscopy. The tenacious nature of the mucous layer was demonstrated by its stability following exposure to formalin and a cationic detergent. Second, the presence of this layer was confirmed on gill tissue which was cryopreserved, followed by freeze-substitution and vapor fixation, and then examined by transmission electron microscopy.  相似文献   

3.
Fixation of mucus for the assessment of biofilms and surface associated pathogens often involves complex and expensive techniques. Rainbow trout killed by an overdose of MS 222 had their gills removed and immersion-fixed gently in buffered glutaraldehyde containing 2% Alcian blue. Control tissues consisted of gills fixed in Alcian blue-free fixative. Trout were also killed and directly immersed in liquid nitrogen and the gills freeze-dried then vapour fixed with osmium tetroxide at −50° C. Following fixation gill tissue was processed for light and electron microscopy. A continuous and intact mucous coat was not detected on tissue fixed by conventional methods but the addition of Alcian blue to the fixative preserved an extensive mucous coat trapped between the lamellae and overlying the epithelia. Electron microscopic examination revealed that mucus preservation with the conventional fixative was poor and intermittent whereas the addition of Alcian blue to the fixative greatly enhanced the preservation of the branchial mucous coat. Mucus appeared as interdispersed flocculant material between the epithelial microridges and formed extensive superficial sheets over the epithelium. Freeze-dried/vapour-fixed gill tissue also provides excellent preservation of the integrity of branchial mucous coats, the mucus appearing as a continuous sheet over the filament and secondary lamellae. However, freeze-dried tissue fails to preserve sufficient cellular integrity for this technique to be useful for light or transmission electron microscopy. The potential for use of glutaraldehyde-Alcian blue fixed-gill tissue diagnostically and in research are discussed.  相似文献   

4.
Summary Scanning electron micrographs of gill tissue from rainbow trout fixed with 50% glutaraldehyde revealed the presence of microridges on surfaces of epithelial cells of the secondary lamellae. These microridges vary in length from 1 to 7 , with a mean height of 0.75 . Calculations show that they increase the total lamellar epithelial surface area approximately 2.5 fold. Mucus secreting cells are present on the body of the filament and on secondary lamellae. Chloride cells are located primarily in the interlamellae filamental epithelium and on the basal area of lamellae. Extensions of the chloride cell epithelium are microvillous in nature and their height is only slightly greater than that of the microridges of typical lamellar epithelial cells. A reduction in number or complete absence of microvilli on chloride cells appeared to be related to degenerative changes in these cells observed in transmission electron micrographs. Non secretory interlamellae filamental epithelial cells have microridges of very attenuated lengths.This research was supported by EPA Grant R-801034, USPHS Training Grant HL-05873, the Mich. Agr. Exp. Sta., Proj. 122 (Journal Article No. 5801), and OWRR Grant A-064. Acknowledgements: The authors wish to express their gratitude to Mrs. J. Mack and Mr. Wm. McAffe for their technical assistance with the electron microscopes.  相似文献   

5.
Variations in the gross morphology and surface architecture of the gill filaments and secondary lamellae of a freshwater catfish (Rita rita) have been investigated using scanning electron microscopy. Heterogeneity of the gill has been correlated with the distribution of lamellar water-flow at different regions of a gill filament. Higher lamellar water flow (cc/pore/cmH2O/sec) was estimated for the middle region of the filaments. The filaments are covered with epithelial cells whose surface is provided with well-developed microridges. The lamellae are generally covered with microvillous epithelial cells. The variations in surface architecture of the gill filaments and secondary lamellae have been correlated with their probable functions.  相似文献   

6.
The integument of the paddlefish (Polyodon spathula) is unusual as a relatively small amount of mucus is produced by epithelial cells that are not modified into regular mucous gland cells. A thick compact epidermis and dermis compensate for the slight amount of mucus secreted. Paddlefish have a variety of scales formed of concentric bony lamellae containing osteocytes. There are five kinds of scales: dorsal and ventral fulcra on the caudal fin, rhomboidal scales on the caudal lobe, horny denticles over the pectoral girdle, calcareous denticles on the trunk, and anchor-shaped plates on the rostrum. Except for the fulcra, the scales are undoubtedly vestigial. The numerous surface pits on the rostrum, head, operculum, and throat are epithelial invaginations which are not connected to lateral line canals. No nerves lead to the pits. The spherical to cuboidal and often ciliated cells at the base of the pits are considered to be aplasic cells of unformed neuromasts.  相似文献   

7.
Summary The gill secondary lamellae are generally covered with epithelial cells whose outer surfaces form numerous microvilli. The surface of the primary lamellae is characterised by microridges. A particular type of surface sculpturing seems to be associated with given cell boundaries.Further evidence for the derivation of the air tube and fans which guard its entrance by modification of the basic gill structure has been obtained from both the gross surface architecture and microstructure of the individual cell surfaces. Secondary lamellae are represented by stubby projections which generally have a biserial arrangement. The outer surfaces of the epithelia overlying the capillaries of these respiratory islets are coated with microvilli as in the secondary lamellae. On the other hand, the relatively smooth-surfaced lanes between groups of respiratory islets have a microridged surface similar to that of the primary gill lamellae.It is suggested that previous estimates of surface area, and consequently diffusing capacities of the air-breathing organ, have been low in view of the increased surface, due to both their gross and microstructure. Estimates for gill surface area may need very little correction as the spaces between the microvilli and microridges are probably filled with mucus under normal conditions.We thank Mr. John Clements for his excellent technical assistance and the Department of Botany, Bristol University for the use of their scanning electron microscope  相似文献   

8.
Odontesthes argentinensis was collected from Mar Chiquita Coastal Lagoon, the Southernmost coastal Atlantic Lagoon of Argentina. The morphology of the gills was analyzed by scanning electron microscopy. The morphology of the superficial structures of the gill filaments and pharyngeal region of the gill arch was discussed and related to their functional aspects. The gills arches are structurally similar to those of other teleosts and bring out the osmoregulatory capacity of this species. The epithelium that covers the surface of the filaments and the pharyngeal region of the gill arch is formed by polygonal pavement cells with conspicuous microridges. These folds in the membrane are not denoted in the epithelium of the respiratory lamellae. Apical crypts of chloride cells are present on the afferent and interlamellar filament surfaces, but are absent elsewhere on the gill arch. The highest density of mucous cells is observed into the gill filament and the pharyngeal region which indicates the existence of a protective strategy of the respiratory lamellae and the pharynx. The epithelium of the gill arches and the rakers is studded with spines. There are taste buds along the whole pharyngeal region that may be associated with their participation in tasting at this zone.  相似文献   

9.
The structure of the mucous layer covering the oesophageal epithelium was analysed by scanning electron microscopy in the eel, Anguilla anguilla . Different fixation procedures were used to conserve the mucus in situ. The mucous layer changes progressively down the oesophagus from a thick dense layer to a very thin fibrous network. The possible roles of these mucous structures in ion absorption are discussed.  相似文献   

10.
To gain insight into the multiple functions of a complex biological structure, the morphology of the pharynx of the larva (ammocoete) of the lamprey Petromyzon marinus was investigated with scanning electron microscopy and histochemistry (PAS and Alcian blue). Features studied include the gills, the parabranchial chambers external to the gills, intrapharyngeal ciliary tracts, the ridged pharyngeal roof, the floor, and the intrapharyngeal taste buds. Significant findings are: (1) All (nonciliated) cells lining these structures are covered with microvilli or microridges. The pattern and packing density of these membrane features vary among different pharyngeal structures. The lumenar membranes of pharyngeal lining cells overlie a mucous prosecretion in the apical cytoplasm, suggesting that the microvilli/ridges on these membranes function to anchor mucus. (2) Patterns of microvilli/ridges on the gill respiratory lamellae differ among ammocoetes of different species. (3) Pharyngeal osmo-regulatory cells (“chloride cells”) could not be identified on the basis of the microvillus/ridge pattern. (4) Two types of ciliary tracts are present within the pharynx. One has tall (x = 13 μm) and densely packed cilia, whereas the other has shorter (x = 7 μm) and less densely packed ones. Because mucus covers both types of tracts their function appears to involve the transport of mucus. (5) Food particles were found on the lateral surfaces of the gill filaments and on the surfaces of the parabranchial chambers. It appears that goblet cells in the epithelia of these regions secrete mucus in which the particles are trapped.  相似文献   

11.
Actin microridges form labyrinth like patterns on superficial epithelial cells across animal species. This highly organized assembly has been implicated in mucus retention and in the mechanical structure of mucosal surfaces, however the mechanisms that regulate actin microridges remain largely unknown. Here we characterize the composition and dynamics of actin microridges on the surface of zebrafish larvae using live imaging. Microridges contain phospho-tyrosine, cortactin and VASP, but not focal adhesion kinase. Time-lapse imaging reveals dynamic changes in the length and branching of microridges in intact animals. Transient perturbation of the microridge pattern occurs before cell division with rapid re-assembly during and after cytokinesis. Microridge assembly is maintained with constitutive activation of Rho or inhibition of myosin II activity. However, expression of dominant negative RhoA or Rac alters microridge organization, with an increase in distance between microridges. Latrunculin A treatment and photoconversion experiments suggest that the F-actin filaments are actively treadmilling in microridges. Accordingly, inhibition of Arp2/3 or PI3K signaling impairs microridge structure and length. Taken together, actin microridges in zebrafish represent a tractable in vivo model to probe pattern formation and dissect Arp2/3-mediated actin dynamics in vivo.  相似文献   

12.
The pseudobranchs of bass, Dicentrarchus labrax L. , and grey mullet, Liza ramada (Risso) ( Mugil capito ), were examined in the scanning electron microscope (S.E.M.). Both pseudobranchs appeared gill-like in situ but the S.E.M. revealed their gross morphology to be different. The bass pseudobranch was a'free' pseudobranch, having separate lamellae along the filaments, with areas of fusion on the leading edges of some lamellae, whilst the mullet pseudobranch was 'semi-free' since the secondary lamellae were fused over a large area and only free along their trailing (opercular) edge where the chloride cells were situated. The surface of the epithelial cells presented different patterns of microridges and microvilli depending on their position in the pseudobranch. Three types of cell opening were found at the cell surface, belonging to chloride cells, mucous cells, and possibly rodlet cells. The S.E.M. observations were correlated with those of transmission electron microscope (T.E.M.) studies.  相似文献   

13.
The general morphology and surface ultrastructure of the gills of adult and larvae medaka (Oryzias latipes) were studied in freshwater and seawater using scanning electron microscopy. The gills of all examined fish were structurally similar to those of other teleosts and consisted of four pairs of arches supporting (i) filaments bearing lamellae and (ii) rakers containing taste buds. Three cell types, specifically pavement cells, mitochondria‐rich cells (MRCs), and mucous cells, constituted the surface layer of the gill epithelium. Several distinctive characteristics of medaka gills were noted, including the presence of regularly distributed outgrowth on the lamellae, enlarged filament tips, the absence of microridges in most pavement cells in the filament and lamellae and the presence of MRCs in the arch at the filament base. A rapid mode of development was recorded in the gills of larval fish. At hatching, the larvae already had four arches with rudimentary filaments, rakers, and taste buds. The rudimentary lamellae appeared within 2 days after hatching. These results suggest the early involvement of larval gills in respiratory and osmoregulation activities. The responses of the macrostructures and microstructures of gills to seawater acclimation were similar in larvae and adult fish and included modification of the apical surface of MRCs, confirming the importance of these cells in osmoregulation. The potential roles of these peculiarities of the macrostructures and microstructures of medaka gills in the major functions of this organ, such as respiration and osmoregulation, are discussed.  相似文献   

14.
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.  相似文献   

15.
PHYSICOCHEMICAL EFFECTS OF ALDEHYDES ON THE HUMAN ERYTHROCYTE   总被引:7,自引:5,他引:2       下载免费PDF全文
The effects of formaldehyde, acetaldehyde, and glutaraldehyde on human red blood cells were investigated. It was found that (a) The surface negative charge of the erythrocytes at pH 7 was increased 10% by glutaraldehyde, but not by the other two aldehydes. (b) The effect of incomplete fixation of the red blood cells was demonstrated by hemoglobin leakage studies The leakage of hemoglobin subsequent to formaldehyde treatment was especially pronounced Acetaldehyde-fixed cells showed some leakage of hemoglobin after an hour of exposure to the fixative, whereas glutaraldehyde-fixed cells showed no hemoglobin leakage. (c) All three aldehydes caused K+ leakage during fixation. The concentrations of K+ in the fixing solutions all reached the same level, but whereas the leakage with glutaraldehyde was immediate, that with formaldehyde was more gradual and that with acetaldehyde reached a steady state only after 24 hr. (d) The effects of the aldehydes on red cell deformability and swelling revealed that glutaraldehyde hardened the cells within 15 min, formaldehyde within 5 hr, while acetaldehyde required at least 24 hr to produce appreciable fixation. (e) The hematocrit changes accompanying the fixation process depended upon cell volume changes and loss of deformability.  相似文献   

16.
In order to characterize the glycoconjugate residues in skin and gills of the adult rainbow trout, the binding pattern of five biotinylated lectins with different carbohydrate specificities was examined. In the skin, mucous cells revealed binding sites for PNA and SBA; filament-containing cells were additionally labelled with Con A. However, the basal cell layer showed no reaction. In the gill, subpopulations of mucous cells reacted with Con A, PNA, SBA and UEA-I. This broader spectrum of glycoconjugates in gill mucous cells compared with the epidermal mucous cells could point to the additional function of gill mucus in ion and osmoregulation. Lectin binding sites were less common in the respiratory epithelial cells of the secondary lamellae than in those of the primary lamellae. Chloride cells revealed mannose, galactose and fucose residues. Immature chloride cells, as indicated by a comparison with Na+/K+ ATPase immunolabelling, reacted with Con A; subpopulations of them reacted with PNA, SBA and UEA-I. The results form the basis for further investigations in which these cell populations can be analysed under different environmental conditions  相似文献   

17.
Glycoproteins (GPs) were visualised histochemically in the secretory cells – the mucous goblet cells (the type A and the type B), the serous goblet cells, the club cells and the epithelial cells in the gill epithelium of Rita rita. The type A mucous goblet cells, the type B mucous goblet cells and the epithelial cells elaborate GPs with oxidizable vicinal diols and GPs with sialic acid residue without O-acyl substitution. In addition, GPs with O-sulphate esters are elaborated by the type A and GPs with O-acyl sugars by the type B mucous goblet cells. GPs are absent in the serous goblet cells and are with oxidizable vicinal diols in low moieties in the club cells. The analysis of the results elucidates interesting differences in the composition and concentration of GPs in the mucus elaborated by the epithelium of the gill arches and the gill rakers; and the gill filaments and the secondary lamellae indicating the potential importance of the glycoproteins at these locations. GPs elaborated on the surfaces of the gill arches and the gill rakers could be associated to assist in feeding activities and on the surfaces of the gill filaments and the secondary lamellae in the respiratory activity.  相似文献   

18.
In order to characterize the glycoconjugate residues in skin and gills of the adult rainbow trout, the binding pattern of five biotinylated lectins with different carbohydrate specificities was examined. In the skin, mucous cells revealed binding sites for PNA and SBA; filament-containing cells were additionally labelled with Con A. However, the basal cell layer showed no reaction. In the gill, subpopulations of mucous cells reacted with Con A, PNA, SBA and UEA-I. This broader spectrum of glycoconjugates in gill mucous cells compared with the epidermal mucous cells could point to the additional function of gill mucus in ion and osmoregulation. Lectin binding sites were less common in the respiratory epithelial cells of the secondary lamellae than in those of the primary lamellae. Chloride cells revealed mannose, galactose and fucose residues. Immature chloride cells, as indicated by a comparison with Na+/K+ ATPase immunolabelling, reacted with Con A; subpopulations of them reacted with PNA, SBA and UEA-I. The results form the basis for further investigations in which these cell populations can be analysed under different environmental conditions This revised version was published online in November 2006 with corrections to the Cover Date.  相似文献   

19.
Some coral species of the genus Porites can produce thick mucous sheets that partially or completely envelope the colony’s surface. This phenomenon has been reported many times, but the cause and ecological significance remains unclear. In this study, sheet production was examined in response to elevated suspended sediment concentrations associated with a large-scale, extended dredging project on a coral reef. Approximately 400 corals at 16 locations situated from 0.2–33 km from the excavation area were examined at fortnightly intervals over the 1.5 year dredging campaign. Mucous sheets were observed on 447 occasions (from 10,600 observations), with average mucous prevalence ranging from 0–10%. Overall 74 ± 5% of the colonies <1.5 km from the dredging produced one or more sheets. High levels of mucous coverage (≥95% of the colony surface) was observed on 68 occasions, and 82% of these occurred at sites close to the dredging. Approximately 50% of colonies produced ≥3 sheets over the monitoring period, and 90% of these were located close to the dredging. In contrast, at distantly located reference sites (>20 km away), mean mucous sheet prevalence was very low (0.2% ± 0.1), no colonies produced more than 1 sheet, and only 1 colony was observed with high mucous coverage. In a laboratory-based experiment, explants of Porites spp. exposed to fine silt also produced mucous sheets (105 sheets recorded in 1100 observations), with nearly 30% of the fragments exposed to repeated sediment deposition events of 10 and 20 mg cm−2 d−1 producing 2 new sheets over the 28 day exposure period. These multiple lines of evidence suggest a close association between mucous sheet formation and sediment load, and that sheet formation and sloughing are an additional mechanism used by massive Porites spp. to clear their surfaces when sediment loads become too high. These results suggest that mucous sheet formation is an effective bioindicator of sediment exposure.  相似文献   

20.
Bats have a very high mass-specific energy demand due to small size and active flight. European bat species are mostly insectivorous and the morphology of the gastrointestinal (GI) tract should be adapted accordingly. This study investigated the general anatomy by histology and the function by analysing carbohydrate distribution in particular of the mucus of the GI tract of the insectivorous bat Pipistrellus pipistrellus. The GI tracts of three individuals were dissected, fixed in formaldehyde, and embedded in paraffin wax. The tissues and cells of the GI tract of P. pipistrellus were analysed by classical (acid alizarin blue, haematoxylin-eosin, and Masson Goldner Trichrome), histochemical (periodic acid-Schiff, Alcian blue at pH 2.5) and lectin histochemical (lectins WGA and HPA) staining procedures. The GI tract of P. pipistrellus is organised into the typical mammalian layers. The short, narrow, and thin-walled esophagus is simple with a folded stratified squamous epithelium without glands but mucous surface cells secreting neutral mucus. The stomach is globular shaped without specialisation. Mucous surface cells produced neutral mucus whereas neck and parietal cells secreted a mixture of neutral and acid mucus. Chief cell surface was positive for N-acetylglucosamine and the cytoplasm for N-acetylgalactosamine residues. The intestine lacked a caecum and appendix. The small intestine was divided into duodenum, jejunum-ileum and ileum-colon. The epithelium consisted of columnar enterocytes and goblet cells. The large intestine is short, only represented by the descending colon-rectum. It lacked villi and the mucosa had only crypts of Lieberkühn. Towards the colon-rectum, goblet cells produced mucus with N-acetylglucosamine residues increasing in acidity except in colon-rectum where acidity was highest in the base of crypts. Along the tube the surface of enterocytes was positive for N-acetylglucosamine and N-acetylgalactosamine. All over the mucus filling the lumen of the GI tract was positive for N-acetylglucosamine and increased in acidity in all parts except of the stomach.In conclusion, the simple GI tract showed an anatomical reduction of tissue enabling for a short retention time and a reduction of the load carried during flight: short GI tract, lack of lymphoid tissue, missing of glands in certain regions, and a distinct pattern of mucus distribution, indicating different physiological functions of these areas. The GI tract of P. pipistrellus was typical for an insectivorous species probably representing the ancestral condition.Key words: Chiroptera, esophagus, glycoconjugates, intestine, lectins, stomach  相似文献   

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