Observations on the relationship between “chloride-type” and “pseudobranch-type” cells in the gills of a fish,Oligocottus maculosus

Summary Chloride cells in gill epithelium of Oligocottus maintained in sea water have a much branched system of agranular cytoplasmic tubules, numerous mitochondria, and a prominent apical crypt. The mitochondria are randomly dispersed and do not show preferential orientation with respect to the tubules.After brief exposure of fish to sea water diluted 1/100 with glass distilled water, the mitochondria and tubules become rearranged into parallel arrays and the apical crypts disappear. The appearance of these cells is similar to that of pseudobranch cells in Fundulus heteroclitus (Copeland and Dalton, 1959).These data suggest that chloride cells and pseudobranch cells represent different adaptive forms of a single cell type and that transformation from the chloride cell configuration to that of pseudobranch cells can be induced by osmotic stress.This work was performed in 1963 when the author was enrolled in the Fine Structure Training program of the Department of Biological Structure, University of Washington. Financial aid was provided by the United States Public Health Service (Grant No. 5T1 GM-136).     

Structure and Function of the Chloride Cell of Fundulus heteroclitus and Other Teleosts

Teleosts, the bony fishes, inhabit both freshwater and seawaterenvironments. Some euryhaline fish, such as Fundulus heteroclitus,alternate between the two milieux several times daily. Regardlessof adaptation, the gills of these animals possess a highly specializedcell type called the chloride cell. This cell contains numerousmitochondria and exhibits a greatly amplified basolateral cellsurface richly endowed with Na,K-ATPase. Recent studies on isolatedopercular epithelia containing chloride cells have demonstratedactive chloride secretion and passive transepithelial sodiummovements, and have established the chloride secretory roleof this cell type in seawater-adapted teleosts. Current modelssuggest that chloride transport occurs via a transcellular route.Seawater chloride cells exist in multicellular units and sharesimple, shallow tight junctions which are thought to be theroute for passive sodium movement. Freshwater chloride cells,whose function remains to be elucidated, are generally describedas existing in a unicellular configuration. However, recentobservations in Fundulus heteroclitus adapted to salinitiesas low as 1% sea water reveal that chloride cells persist inmulticellular complexes with apical crypts. Strikingly, tightjunctions between chloride cells in this freshwater environmentare deep     

Ultrastructure of chloride cells in young adults of the anadromous sea lamprey, Petromyzon marinus L., in fresh water and during adaptation to sea water.

The chloride cells in the interlamellar areas of the gills of young adult, anadromous sea lampreys, Petromyzon marinus L., captured in fresh water undergo structural modification during the adaptation of these animals to sea water. In fresh water the chloride cells are partially overlapped by mucus-secreting superficial cells and contain an extensive reticulum of cytoplasmic tubules, which are confluent with both lateral and basal plasma membranes, numerous mitochondria, a Golgi complex of moderate size, and numerous apical vesicles. Adaptation to sea water results in a retraction of the superficial cells, exposing the entire apical surface of the chloride cells, and a proliferation of both cytoplasmic tubules and mitochondria. Extensive enlargement of the Golgi complex in the chloride cells of these animals suggests the involvement of this organelle in the proliferation of cytoplasmic tubules. The extracellular tracer, ruthenium red, enters the tubules from the lateral or basal intercellular spaces in both freshwater- and seawater-adapted animals but never enters either tubules or vesicles from the apical surfaces, indicating that these are not confluent. The presence of dividing basal cells and newly-forming chloride cells, combined with evidence of degeneration of chloride cells, suggests that there is a turnover of this cell type. Both superficial and basal cells are phagocytic and involved in heterophagy of degenerating chloride cells. This phenomenon occurs in both fresh water and sea water indicating that the chloride cells may be functional in both environments.     

Mitochondria-rich (chloride) cells in the gill epithelia from four species of stenohaline fresh water teleosts

Summary The mitochondria-rich (chloride) cells have been found to be present in the gill epithelia of four species of stenohaline fresh water teleosts. The cytoplasm of these chloride cells contains an extensive network of cytoplasmic tubules which communicate with intercellular spaces bordering the lateral and basal cell surfaces. Numerous vesicles with fairly electron-dense interiors are also present in the apical cytoplasm of chloride cells. The apical surface of a chloride cell forms an apical pit, but the lumen of the pit does not appear to be in continuity with the interior of the apical vesicles and tubules inside the cell.When Carassius auratus were kept in 100, 200, 300, and 400 mOsm-diluted sea water for a month, no appreciable changes occurred in the number and fine structure of the chloride cells, except for a dilation of the apical vesicles and a slight decrease in diameter of the cytoplasmic tubules in these cells in the fishes kept in 300 and 400 mOsm.These results suggest that chloride cells may be a rather common occurrence in the gill epithelia of stenohaline fresh water teleosts, and may function in ion-transport in these fishes in fresh water environments.     

Isoenzymes de l'anhydrase carbonique d'un poisson euryhalin: Variations en relation avec l'osmoregulation

A chemical study of carbonic anhydrase (EC 4.2.1.1) from the red blood cells and the gills of an euryhaline fish (Anguilla anguilla) is presented. Animals adapted to fresh water were compared to those adapted to sea water.The physicochemical constants of the various molecular forms isolated by chromatograhy and isoelectric focusing were determined; isoeleletric pH, molecular weight, and the K_m and V/E of the enzyme dehydration activity were compared.In both red cells and gills of fish adapted in either media various forms were isolated, characterized by different enzymatic kinetics (high- and low-activity forms) but having the same molecular weight (27 250). Some isoenzymes isolated from the gills differed significantly from those isolated from the red cells.Adaptation to fresh water or sea water is accompanied by modifications in the distribution of yhe isoenzymes in both red cells and gills: adaptation to sea water is characterized by a shift of the molecular forms towards an isoelectric pH higher than pH = 6.The role of these enzymes is discussed under both a physiological and biochemical point of view in relation to the electrolyte exchange across fish gill. The origin of the different molecular forms of the carbonic anhydrase is discussed in relation to the prevailing theories on this subject.     

Die Chloridzellen des Stichlings

Zusammenfassung Dreistachlige Stichlinge (Gasterosteus aculeatus) aus Süßwasserbiotopen wurden in mehreren Versuchsgruppen allmählich an Meersalzlösungen steigender bzw. fallender Konzentration adaptiert. Dabei stellte sich heraus, daß diese euryhaline Fischart Salzkonzentrationen zwischen 1 mg-% und 5,6% tolerieren kann. Der letzte Wert bedeutet das 1,6fache der durchschnittlichen Meerwasserkonzentration. Stichlinge aus verschiedenen salzreichen und salzarmen Adaptationsstufen dienten als Ausgangsmaterial zur elektronenmikroskopischen Untersuchung der Chloridzellen.Die Feinstruktur der Chloridzellen zeigt in Abhängigkeit vom Salzgehalt des Mediums typische Veränderungen. Bei Süßwasserstichlingen ist die apikale Höhle septiert und dadurch die resorptive apikale Zellmembranoberfläche vergrößert. Bei Meerwasserstichlingen scheint das endoplasmatische Reticulum der Chloridzellen vermehrt zu sein; ihr Chondriom nimmt 50% des Cytoplasmavolumens ein, bei den Chloridzellen der Süßwassertiere hingegen nur 20%. Im Bereich letaler Salzarmut und letalen Salzreichtums treten bei den Chloridzellen Strukturschädigungen auf.Durch histochemische Ionenfällung konnte in der Mucoidschicht der apikalen Höhle ein hoher Gehalt an Na⁺ und Cl^– nachgewiesen werden. Die Mucoidschicht dieser Zellen füngiert demnach bei Süßwassertieren als akkumulativer Ionenfänger, bei Meerwassertieren möglicherweise als Ionenpuffer.Die Feinstruktur der Chloridzellen, ihre Veränderungen in Abhängigkeit von dem äußeren Salzgehalt, die Schädigungen bei letalen Konzentrationen sowie insbesondere die Ergebnisse der histochemischen Ionenfällung können insgesamt als Beweis der osmoregulatorischen Funktion dieser Zellen gelten.

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The Chloride Cells of the Stickleback

Summary Several groups of the stickleback (Gasterosteus aculeatus) collected from freshwater were gradually adapted to sea salt solutions of increasing resp. decreasing concentrations. As shown by these adaption experiments, this euryhaline teleost species is able to tolerate salt concentrations in the range of 1mg-% to 5,6%. The last value corresponds to 1,6 times of the average salt concentration of sea water. Specimen adapted to minimum, intermediate and maximum salt concentration were chosen for electron microscopical investigation of the chloride cells.Depending on the external salt concentration the fine structure of these cells shows typical alterations. In fresh water specimen, the apical cavity of the chloride cells is septate and consequently the resorptive apical cell membrane surface is enlarged. In sea water specimens the endoplasmic reticulum seems to be more developed; the mitochondria take about 50% of the cytoplasm volume, whereas they take only 20% in fresh water animals. The chloride cell fine structure of those animals which had been brought to the upper or lower limit of the tolerable salt concentration is damaged.The mucoid layer of the apical cavity in animals from both salt-rich and nearly salt-free medium has a high content of Na⁺ and Cl^–, as detected by histochemical ion precipitation methods. Therefore in fresh water specimens the mucoid layer must be involved in adsorbing and in accumulation ions from the external medium, in sea water specimens its function seems to be something like an ion buffer.From these findings there is no doubt that the osmoregulatory function of the teleost gills is based on the chloride cells.

     

Patterns in the utilization of the intertidal salt marsh by larvae and juveniles of Fundulus heteroclitus (Linnaeus) and Fundulus luciae (Baird)

Larval and juvenile fishes were collected at low tide from the surface of an intertidal salt marsh on Sapelo Island, Georgia every 6 days from 25 May through 20 December 1982. Larval fishes were present in shallow puddles of tidal water on the marsh from the beginning of the sampling period until the end of October. Juveniles were present throughout the year. Fundulus heteroclitus (Linnaeus) and F. luciae (Baird) accounted for 96.3% (67.0% and 29.3%, respectively) of the 4355 fishes collected. These fishes exhibited synchronous, temporal pulses in larval abundance, suggesting that reproduction was discontinuous and controlled by the same factor(s) in both species. Larval F. heteroclitus inhabited aquatic micro-habitats near the upland edge of the intertidal marsh, but as the larvae grew to juvenile size (≈ 10 mm standard length) they moved to lower elevations near a tidal creek. Large juveniles and adults of F. heteroclitus infrequently occurred in the samples, presumably because they leave the intertidal marsh as the tide ebbs. F. luciae were rarely found in low marsh areas, but all age classes (including adults) occurred at higher elevations, supporting previous suggestions that this species prefers high marsh habitats. The vegetated, intertidal salt marsh appears to be the principal nursery habitat for both of these cyprinodontid species.     

The ClC-3 chloride channel and osmoregulation in the European Sea Bass, Dicentrarchus labrax

Dicentrarchus labrax migrates between sea (SW), brackish and fresh water (FW) where chloride concentrations and requirements for chloride handling change: in FW, fish absorb chloride and restrict renal losses; in SW, they excrete chloride. In this study, the expression and localization of ClC-3 and Na⁺/K⁺-ATPase (NKA) were studied in fish adapted to SW, or exposed to FW from 10 min to 30 days. In gills, NKA-α1 subunit expression transiently increased from 10 min and reached a stabilized intermediate expression level after 24 h in FW. ClC-3 co-localized with NKA in the basolateral membrane of mitochondria-rich cells (MRCs) at all conditions. The intensity of MRC ClC-3 immunostaining was significantly higher (by 50 %) 1 h after the transfer to FW, whereas the branchial ClC-3 protein expression was 30 % higher 7 days after the transfer as compared to SW. This is consistent with the increased number of immunopositive MRCs (immunostained for NKA and ClC-3). However, the ClC-3 mRNA expression was significantly lower in FW gills. In the kidney, after FW transfer, a transient decrease in NKA-α1 subunit expression was followed by significantly higher stable levels from 24 h. The low ClC-3 protein expression detected at both salinities was not observed by immunocytochemistry in the SW kidney; ClC-3 was localized in the basal membrane of the collecting ducts and tubules 7 and 30 days after transfer to FW. Renal ClC-3 mRNA expression, however, seemed higher in SW than in FW. The potential role of this chloride channel ClC-3 in osmoregulatory and osmosensing mechanisms is discussed.     

Overwintering habitat selection by the mummichog, Fundulus Heteroclitus, in a Cape Cod (USA) salt marsh

This study tracked the seasonal distribution and winter habitat selection of the mummichog, Fundulus heteroclitus (Linnaeus), in a Cape Cod, Massachusetts salt marsh. Fish (mean size = 43.1 mm total length, range = 10–93 mm) were collected with a 1 m² throw trap and by excavating sediments. In fall, F. heteroclitus began migrating upstream in creeks and eventually moved into upstream pools where they remained throughout winter. F. heteroclitus burrowed into the sediments of these pools at a density of 0.5 fish m^–2, but was not found burrowed in the sediments of downstream pools or any creeks. Sediments in upstream pools were composed of a higher proportion of fine-grained particles and organic content than other marsh pools and creeks, and winter temperatures in upstream pool sediments remained above 1 °C. Temperatures in the water column and sediments of downstream pools regularly dropped below –1.8 °C, exceeding the lethal limit for F. heteroclitus. These results support other recent work showing that F. heteroclitus migrates upstream in salt marshes in fall and overwinters in salt marsh pools. Moreover, this study demonstrates that F. heteroclitus does not utilize all available pools as overwintering habitat but apparently selects pools with sediments that offer a thermal refuge from lethal winter temperatures.     

Changes in osmotic water permeability of the eel gills during seawater and freshwater adaptation

Summary Changes in osmotic water permeability of the isolated gills of the Japanese eel,Anguilla japonica were studied during transfer to seawater or to fresh water. The water permeability increased gradually during the course of seawater transfer and attained a maximal level after 2 weeks. The water permeability of the freshwater eel gills was reduced when calcium ions were added to the incubation medium at a concentration of 1 mM, where-as no effect of the ion was observed on the gills of the seawater-adapted eel even at a higher concentration (10 mM). In contrast to seawater transfer, the water permeability decreased to a low level almost immediately (3 h) after transfer from seawater to fresh water. The acute reduction of the water permeability was also seen in the gills of the hypophysectomized eel after transfer to fresh water.The gradual increase in the gill water permeability during seawater transfer is correlated with an increase in the number of chloride cells. In scanning electron microscopy, chloride cells of seawater-adapted eel gills exhibit a pit-like structure, which was larger than in the freshwater eel. On transfer from seawater to fresh water, the pit diameter became smaller within 6 h. Hypophysectomy did not affect the change in gill surface structures during transfer to fresh water. The junctions between the chloride cells of seawater eel gills are reported to be of the leaky type. The parallel change in osmotic water permeability and in pit size of the chloride cells during seawater or freshwater transfer or after hypophysectomy suggests that these cells could provide a major route of water as well as ion movement.This paper is a portion of a thesis presented to Hokkaido University by t. Ogasawara in partial fulfilment of the requirements for Doctor of Fisheries     

The demonstration of chloride ions in the “chloride cells” of the gills of eels (Anguilla anguilla L.) adapted to sea water

Summary The ultrastructure of chloride cells in the gills of eels kept in artificial sea water and of a control animal kept in fresh water was studied. In addition to glutaraldehydeosmium tetroxide and simple osmium tetroxide fixation, a special method for the demonstration of chloride ions was used (Komnick, 1962, 1963). Based on the principle of silver chloride precipitation in the presence of chloride ions, the procedure showed positive results in the chloride cells of eels adapted to sea water. Smooth-surfaced tubules of the endoplasmic reticulum contained a material of medium to strong electron density, that was often in communication with the plasma membrane. The same material, always of very high density, was present in the intercellular spaces, thus forming conspicuous lines around cells. The silver precipitate was found very often in large quantities in the pits of chloride cells, having thereby the aspect of a secretory product. However, a direct communication between the system of endoplasmic reticulum tubules containing the silver reaction product with the above-mentioned masses of silver chloride was not demonstrated. Schultz (1958) first published an electron micrograph of these cells. Two features, numerous mitochondria and highly developed agranular endoplasmic reticulum, affirmed by all later investigators, have remained the most important criteria for the identification of chloride cells (Kessel and Beams, 1962; Philpott, 1962; Philpott and Copeland, 1963; Rhodin, 1964; Henrikson and Matoltsy, 1968). Kessel and Beams (1962) and Philpott and Copeland (1963) demonstrated pits, or apical cavities (Oberg, 1967) filled with an amorphous granular substance of medium electron density. (Threadgold and Houston, 1964). These cavities are found in animals adapted to sea water only, and correspond to the excretory vesicles described earlier by Copeland (1948). Philpott and Copeland demonstrated numerous vesicles and tubules in the cytoplasm surrounding the apical cavities and opening into them, possibly contributing to their granular material. Several authors attempted to demonstrate a chloride excretory function of chloride cells with the aid of some histochemical reactions. Copeland (1948), Datta Munshi (1964) and Philpott (1966) used silver techniques     

The distribution of intracellular ions in the avian salt gland

To investigate the mechanism of salt secretion in the avian salt gland, we used quantitative electron probe microanalysis to measure the intracellular elemental concentrations in dry cryosections of unspecialized and partially specialized secretory epithelial cells from fresh water- and salt water-adapted ducklings, respectively. In conjunction with this, human and duckling erythrocytes were also analyzed, since these provided the experimental basis for using in situ erythrocytes as standards for determining the local water content of epithelia from the analysis of dried cryosections. The microprobe results from both types of erythrocytes compared favorably with chemical determinations of elemental concentrations. The nucleated avian erythrocytes, whose wet-weight elemental concentrations were determined by a compartmental analysis that required neither a peripheral standard nor a measure of the local mass, revealed a marked accumulation of P and K in the nucleus (388 and 190 mmol/kg wet wt, respectively) relative to the cytoplasm (67 and 85 mmol/kg wet wt). In both developmental states of the epithelial cells, the nucleus and apical cytoplasm had essentially similar and unremarkable concentrations of Na (76 and 83 mmol/kg dry wt, respectively, in the adapted cells vs. 72 and 81 mmol/kg dry wt in the control cells) and K (602 and 423 mmol/kg dry wt vs. 451 and 442 mmol/kg dry wt). Chloride, however, which was in general rather high, was significantly depressed in the apical cytoplasm of adapted cells only (164 and 124 mmol/kg dry wt in the nucleus and cytoplasm, respectively, of adapted cells (P less than 0.05) vs. 138 and 157 mmol/kg dry wt for control cells (P less than 0.05). Cation concentrations (Na + K) were elevated approximately 15% in the basal regions of adapted cells as compared with apical cytoplasm. When tissue water variations are accounted for, the results suggest that: (a) an active, energy-requiring process is responsible for chloride accumulation in this cell; (b) the apical membrane is a regulatory site for secretion; and (c) there are regional distinctions in the distribution of ions and water, particularly in the salt water- adapted cell. These conclusions are consistent with active chloride transport as the basis for salt secretion in this tissue.     

Effect of different salinities on the coniform chloride cells of mayfly larvae

The larvae of Callibaetis coloradensis can tolerate a fairly wide range of salinities at hypotonic concentrations. However, they are more sensitive to increasing than to decreasing salt concentrations. Exposure to isotonic concentration results in profound degenerations of the chloride cells within 1 day. Long-term adaptation to diluted fresh water causes a significant increase in the number of chloride cells, whereas the gradual concentration of fresh water to finally 120 mM sodium chloride within a period of 15 days leads to approximately 50 per cent mortality and significantly reduces the number of chloride cells in the survivors. The same reciprocal relation between the number of chloride cells and the external salinity was found in larvae of C. floridans collected from fresh- and brackish-water habitats. These results suggest that the adaptive behaviour of the chloride cells is correlated with the osmoregulatory situation and enables these animals to live in habitats of different salinities.     

Anomalous tolerance to low pH in the estuarine killifish Fundulus heteroclitus

1. The euryhaline fish Fundulus heteroclitus has an incipient lethal pH between 3.75 and 4.0 in fresh water.2. Fish exposed to pH 3.5 in sea water or fresh water died in about 3 hr, and had greatly elevated or depressed body sodium concentrations, respectively. The direction and degree of change in body sodium level depended on the sodium diffusion gradient between the environment and the fish. This is the first time that the death of fish in sea water at low pH has been shown to be associated with hypernatremia.3. Yet, sodium fluxes during the first hour of exposure to pH 3.5 in water of 3.5 or 35 ppt salinity were not different from controls, and body and plasma sodium concentration did not change during 2hr exposure to pH 3.5. This initial insensitivity of gill sodium regulation to blockage by low pH is quite different from the response of previously studied freshwater fish.4. The degree of acid tolerance displayed by F. heteroclitus is surprising considering its estuarine habits. This paradoxical tolerance appears to be a secondary consequence of its ability to adjust sodium balance in relation to rapid changes in salinity.     

Habitat use, movement, and growth of young-of-the-year Fundulus spp. in southern New Jersey salt marshes: Comparisons based on tag/recapture

We examined habitat use, movement, and growth of young-of-the-year (YOY) Fundulus heteroclitus and Fundulus luciae with a tag/recapture experiment in tide-dominated salt marshes to determine if movements from Spartina marsh surface can account for the occurrence of larger, older individuals in other habitats. Evaluation of the tagging techniques in laboratory experiments with YOY F. heteroclitus (15-35 mm TL) found that coded wire tags were retained at least up to 77 days. The high rates of recapture in the field also indicate that the tagging approach generally worked well. Of a total of 5748 YOY F. heteroclitus (14-40 mm TL) and 133 YOY F. luciae (17-40 mm TL) tagged, 56.0% and 74.4% were recaptured, respectively. Most (44%) YOY F. heteroclitus recaptured occurred at or near (0-5 m) the release site, but some were captured up to 299 m away up to 166 days after tagging. By comparison, movement of F. luciae was very limited, with 99% of recaptures occurring at the exact site of release after up to 66 days at liberty. These different movement patterns by YOY Fundulus indicate that species-specific behavior plays an important role in habitat selection. In addition, it appears that dispersal of YOY F. heteroclitus can help to explain the occurrence of larger individuals of this species in Phragmites-dominated marshes even though there is little evidence of use of this habitat by small YOY.     

Fine structure of chloride cells in freshwater- and seawater-adapted Oreochromis niloticus (Linnaeus) and Oreochromis mossambicus (Peters)

Ultrastructural features of branchial chloride cells in Oreochromis niloticus (Linnaeus) and O. mossambicus (Peters) adapted to both fresh water and sea water were compdred. In freshwater adapted fish of both species chloride cells showed similar morphological features. Multicellular complexes made of a mature chloride cell and one or more accessory cells sharing a single apical crypt have been observed. Whereas high percentages of 0. mossambicus survived at maximum salinity only a few individuals of 0. niloticus showed the capacity to adapt to sea water. In the seawater-adapted individuals of 0. niloficus and 0. rnossambicus the chloride cells showed a two- and three-fold increase in sizei. respectively. Most chloride cells are organized in large multi-cellular complcxcs with apical interdigitations of accessory cells and 'leaky junctions'. These results indicate that the difference in euryhalinity of the species studied is related to functional rather than structural differences.     

The surface epithelium of teleostean fish gills. Cellular and junctional adaptations of the chloride cell in relation to salt adaptation

Various species of teleostean fishes were adapted to fresh or salt water and their gill surface epithelium was examined using several techniques of electron microscopy. In both fresh and salt water the branchial epithelium is mostly covered by flat respiratory cells. They are characterized by unusual outer membrane fracture faces containing intramembranous particles and pits in various stages of ordered aggregation. Freeze fracture studies showed that the tight junctions between respiratory cells are made of several interconnecting strands, probably representing high resistance junctions. The organization of intramembranous elements and the morphological characteristics of the junctions do not vary in relation to the external salinity. Towards the base of the secondary gill lamellae, the layer of respiratory cells is interrupted by mitochondria-rich cells ("chloride cells"), also linked to respiratory cells by multistranded junctions. There is a fundamental reorganization of the chloride cells associated with salt water adaptation. In salt water young adjacent chloride cells send interdigitations into preexisting chloride cells. The apex of the seawater chloride cell is therefore part of a mosaic of sister cells linked to surrounding respiratory cells by multistranded junctions. The chloride cells are linked to each other by shallow junctions made of only one strand and permeable to lanthanum. It is therefore suggested that salt water adaptation triggers a cellular reorganization of the epithelium in such a way that leaky junctions (a low resistance pathway) appear at the apex of the chloride cells. Chloride cells are characterized by an extensive tubular reticulum which is an extension of the basolateral plasma membrane. It is made of repeating units and is the site of numerous ion pumps. The presence of shallow junctions in sea water-adapted fish makes it possible for the reticulum to contact the external milieu. In contrast in the freshwater-adapted fish the chloride cell's tubular reticulum is separated by deep apical junctions from the external environment. Based on these observations we discuss how solutes could transfer across the epithelium.     

Scanning and transmission electron microscopy of the squamose gill-filament epithelium from fresh- and seawater adaptedTilapia

Synopsis The architecture of the gill structure of variousTilapia species was studied in relation to their adaptability to hypersaline media. Using SEM and EM, it was shown that the squamose epithelial cells of the gills have species-typical patterns of ridges on their outer surfaces. These have previously been misinterpreted by other authors as microvilli or stereocillia. The ridges are more dense and better developed in euryhaline species, likeT. zillii, and less so in stenohaline species likeSarotherodon niloticus. Comparing freshwater and seawater-adapted individuals ofT. zillii, S. niloticus, S. galflaeus, andTristramella sacra, it was shown that in fresh water the surface cells are slightly swollen, extending over the openings of the chloride cells. During adaptation to sea water, these ridges become higher and denser and the cell surface shrinks, exposing the underlying orifices of the apical crypts of the chloride cells. The more euryhaline the species, the less change there is in the ridge pattern of the cells during passage from fresh to sea water. This evidence implicates the gill epithelium, together with the chloride cells, in the process of osmoregulation.     

Localisation autoradiographique des ions Cl− et Na+ dans les cellules à chlorure de la branchie d'anguille (Anguilla anguilla L.) adaptée à l'eau de mer

Résumé Nous avons montré par une technique autoradiographique que les ions Cl^– et Na⁺ sont concentrés dans les cellules à chlorure de la branchie d'anguille adaptée à l'eau de mer. La signification de cette accumulation ionique plus marquée vers le pôle apical de ces cellules a été discutée par rapport à l'excrétion branchiale de ces ions en eau de mer.

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Autoradiographic localization of Cl^– and Na⁺ ions in the chloride cells of sea water adapted eel (Anguilla anguilla L.) gills

Summary With an autoradiographic technique Cl^– and Na⁺ ions have been shown to be localized in the chloride cells of sea water eel gills. The significance of this accumulation, more marked towards the apical pole of these cells, is discussed with regard to branchial excretion of these ions in sea water.

     

The Role of Fundulus heteroclitus in Salt Marsh Trophic Dynamics

The common mummichog, Fundulus heteroclitus, functions as bothpredator and prey in the trophic structure of east coast tidalmarshes. Although mummichogs are generally considered importantto energy transformations within marshes, few studies have convincinglydemonstrated that predation by F. heteroclitus affects the abundanceof salt marsh benthic invertebrates. Thus far investigationsof this type have dealt only with the direct effects of adultmummichogs. The results of recent experiments have suggestedthat by controlling smaller predators, mummichogs may indirectlyhave a positive effect on the densities of some infaunal marshinvertebrates. Our current knowledge of larval and juvenilemummichogs in their natural habitat is minimal. Unlike the adults,which can utilize the intertidal zone only when it is flooded,the young remain on the marsh even at low tide, inhabiting shallowpuddles of residual tidal water that form between clumps ofvegetation and around fiddler crab (Uca sp.) burrows. The importanceof F. heteroclitus in salt marsh communities will remain incompletelyunderstood unless future studies consider the role of larvaland juvenile mummichogs. Although many species of fishes andwading birds feed on mummichogs, the blue crab (Callinectessapidus) is probably the major predator of adult F. heteroclitusin the intertidal salt marsh. Predation by adult mummichogsand xanthid crabs (e.g., Eurytium limosum) may contribute tothe high mortality of larval and juvenile Fundulus.