Gill structure of a fish from an alkaline lake: effect of short-term exposure to neutral conditions

1995 ◽  
Vol 73 (6) ◽  
pp. 1170-1181 ◽  
Author(s):  
Pierre Laurent ◽  
J. N. Maina ◽  
Harold L. Bergman ◽  
Annie Narahara ◽  
Patrick J. Walsh ◽  
...  
Keyword(s):  
Sea Water ◽  
Morphological Changes ◽  
Chloride Cells ◽  
Pavement Cells ◽  
Lake Magadi ◽  
Accessory Cells ◽  
Short Term Exposure ◽  
Gill Structure ◽  
Morphology And Morphometry ◽  
Neutral Conditions

The morphology and morphometry of the gills of Oreochromis alcalicus grahami, a unique ureogenic teleost that lives in the alkaline environment of Lake Magadi, Kenya (pH 10, [Formula: see text], temperature 30 – 40 °C) were examined by transmission electron, scanning electron and light microscopy. Fish were examined in normal Lake Magadi water and 2 – 3 or 24 h after transfer to Lake Magadi water neutralized to pH 7 with HCl (i.e., [Formula: see text] replaced with Cl−), a treatment that caused severe reductions in urea excretion and O2 uptake, internal acidosis, and ionoregulatory disturbance. In Lake Magadi water, the organization of the filament epithelium of the gill was similar to that of sea water teleosts. Indeed, chloride cells were located at the bottom of pits bordered by overlying pavement cells and flanked by typical accessory cells. Total numbers of chloride cells remained unchanged after transfer to pH 7, but after 2 – 3 h, many were covered by pavement cells, restricting their communication with the external milieu. At 24 h, this trend was reversed, an observation indicative of a reactivation of chloride cells. Mucous cells were located at maximum density on the trailing edge of the filament; most of them were empty after 24 h at pH 7. The harmonic mean thickness of the lamellar epithelium (blood-to-water diffusion pathway) was very small and not altered by acute or longer term exposure to pH 7. A model of alterations in ion and acid – base transport accompanying the morphological changes is presented.

Shortening of Branchial Tight Junction Acid-Exposed Rainbow Trout (Oncorhynchus mykiss)

1991 ◽  
Vol 48 (10) ◽  
pp. 2028-2033 ◽  
Author(s):  
J. Freda ◽  
D. A. Sanchez ◽  
H. L. Bergman
Keyword(s):  
Rainbow Trout ◽  
Oncorhynchus Mykiss ◽  
Tight Junctions ◽  
Relative Magnitude ◽  
Chloride Cells ◽  
Pavement Cells ◽  
Rainbow Trout Oncorhynchus Mykiss ◽  
Plasma Electrolytes ◽  
Gill Structure ◽  
Electrolyte Loss

The objective of this study was to investigate possible sites for Na+ loss in fish exposed to low environmental pH. In rainbow trout (Oncorhynchus mykiss) exposed to pH 4.0 for 1 h, a net loss of Na+ was stimulated, and changes in gill structure occurred. In addition to epithelial lifting and necrosis in the gills of acid-exposed fish, tight junctions between pavement epithelial cells and chloride cells decreased in length by 25% whereas tight junctions between adjacent pavement cells did not significantly change. In a second experiment where fish were moved from pH 4.0 or 3.5 water to pH 6.5 water, we observed that Na+ loss declined immediately and approached control levels. The reversible nature of the stimulation of Na+ loss indicates that the site of Na+ loss in the fish gill can be reversibly opened and closed, which is consistent with the known properties of tight junctions. We hypothesize that the opening of tight junctions contributes to the loss of plasma electrolytes at low environmental pH. However, the relative magnitude of electrolyte loss through the tight junctions remains unknown.

Ionoregulatory strategies and the role of urea in the Magadi tilapia (Alcolapia grahami)

2002 ◽  
Vol 80 (3) ◽  
pp. 503-515 ◽  
Author(s):  
Chris M Wood ◽  
Paul Wilson ◽  
Harold L Bergman ◽  
Annie N Bergman ◽  
Pierre Laurent ◽  
...  
Keyword(s):  
Lake Water ◽  
Plasma Cortisol ◽  
Sea Water ◽  
Whole Body ◽  
Chloride Cells ◽  
Water Type ◽  
Plasma Ions ◽  
Lake Magadi ◽  
Simultaneous Control ◽  
Alcolapia Grahami

The unique ureotelic tilapia Alcolapia grahami lives in the highly alkaline and saline waters of Lake Magadi, Kenya (pH ~10.0, alkalinity ~380 mmol·L–1, Na+ ~350 mmol·L–1, Cl– ~110 mmol·L–1, osmolality ~580 mosmol·kg–1). In 100% lake water, the Magadi tilapia maintained plasma Na+, Cl–, and osmolality at levels typical of marine teleosts and drank the medium at 8.01 ± 1.29 mL·kg–1·h–1. Gill chloride cells were predominantly of the sea water type (recessed, with apical pits) but a few freshwater-type chloride cells (surficial, with flat apical exposure) were also present. Whole-body Na+ and Cl– concentrations were relatively high and exhibited larger relative changes in response to salinity transfers than did plasma ions. All fish succumbed upon acute transfer to 1% lake water, but tolerated acute transfer to 10% lake water well, and gradual long-term acclimation to both 10 and 1% lake water without change in plasma cortisol. Plasma osmolytes were here maintained at levels typical of freshwater teleosts. Curiously, drinking continued at the same rate in fish adapted to 1% lake water, but chloride cells were now exclusively of the freshwater type. Significant mortality and elevated cortisol occurred after acute transfer to 200% lake water. However, the fish survived well during gradual adaptation to 200% lake water, although plasma cortisol remained chronically elevated. Urea levels accounted for only 2–3% of internal osmolality in 100% lake water but responded to a greater extent than plasma ions during exposure to 10 and 200% lake water, decreasing by 28–42% in the former and increasing by over 500% in the latter relative to simultaneous-control values. Urea thereby played a small but significant role (up to 8% of internal osmolality) in osmoregulation.

scholarly journals Redistribution of immunofluorescence of CFTR anion channel and NKCC cotransporter in chloride cells during adaptation of the killifish Fundulus heteroclitus to sea water

2002 ◽  
Vol 205 (9) ◽  
pp. 1265-1273 ◽  
Author(s):  
W. S. Marshall ◽  
E. M. Lynch ◽  
R. R. F. Cozzi
Keyword(s):  
Fundulus Heteroclitus ◽  
Apical Membrane ◽  
Sea Water ◽  
Basolateral Membrane ◽  
Anion Channel ◽  
Chloride Cells ◽  
Transmembrane Conductance Regulator ◽  
Pavement Cells ◽  
Cell Complexes ◽  
C Terminus

SUMMARY Cellular distribution of cystic fibrosis transmembrane conductance regulator (CFTR) immunofluorescence was detected by monoclonal antibody directed to the C terminus of killifish CFTR (kfCFTR) in chloride cells of fresh water (FW) adapted fish and animals transferred to sea water (SW) for 24h, 48h and 14+ days. Confocal microscopy allowed localization within mitochondria-rich (MR) cells to be determined as superficial (i.e. in the apical membrane) or deeper within the cytoplasm of the cells. In FW, 90 % of MR cells had diffuse kfCFTR immunofluorescence in the central part of the cytosol, with only 8.1 % having apical kfCFTR, which was 6.6±0.54 μm below the microridges of surrounding pavement cells. Curiously, FW but not SW pavement cells also had positive immunofluorescence to kfCFTR. After 24h in SW, a time when kfCFTR expression is elevated, a condensed punctate immunofluorescence appeared among 18.8 % of MR cells, 13.4±0.66 μm(mean ± S.E.M.) below the surface of the cells. By 48h, a majority(76.3 %) of MR cells had punctate kfCFTR distribution and the distance from the surface was less (7.8±0.2 μm), a distribution approaching the SW-acclimated condition (i.e. all MR cells showing kfCFTR immunofluorescence,6.1±0.04 μm below the surface). In contrast, NKCC immunofluorescence was condensed and localized in lateral parts of MR cell complexes in FW animals and then redistributed to the whole basal cytoplasm after acclimation to SW. CFTR, the anion channel responsible for Cl- secretion in marine teleosts, redistributes in MR cells during SW acclimation by condensation of a diffuse distribution below the apical crypt, followed by translocation and insertion in the apical membrane. NKCC, the cotransporter that translocates Cl- across the basolateral membrane, moves from an eccentric cytosolic location in FW to a diffuse basolateral localization in SW chloride cells.

Accessory cells in teleost branchial epithelium

1980 ◽  
Vol 238 (3) ◽  
pp. R199-R206 ◽  
Author(s):  
S. R. Hootman ◽  
C. W. Philpott
Keyword(s):  
Chloride Cell ◽  
Chloride Cells ◽  
Reaction Products ◽  
Cell Type ◽  
Pavement Cells ◽  
Lagodon Rhomboides ◽  
Accessory Cells ◽  
A Cell ◽  
Branchial Epithelium ◽  
Atpase Localization

Correlated morphological and cytochemical investigations of the branchial epithelium of the pinfish, Lagodon rhomboides, have revealed a cell type that is invariably associated with chloride cells. These cells, termed “accessory cells,” have been described previously in the teleost pseudobranch (Dunel and Laurent. J. Microsc. Biol. Cell 16:53-74, 1973) but not in the gill proper. Accessory cells are more numerous in pinfish adapted to seawater than to 33% seawater, and in the former participate with chloride cells in the formation of apical crypts. Although accessory cells are much smaller than chloride cells, they possess numerous mitochondria and display an abbreviated labyrinth of plasma membrane-derived tubules. The labyrinth membranes of accessory cells are essentially unreactive, however, when processed for Na-K-ATPase localization by K-nitrophenylphosphatase cytochemistry, whereas chloride cell membranes exhibit copious, ouabain-sensitive reaction products. The zonulae occludentes between accessory cells and chloride cells also appear to be less extensive than those between either of these cells and the flanking pavement cells. These features suggest that accessory cells represent a population of partially differentiated chloride cells.

In vivo sequential changes in chloride cell morphology in the yolk-sac membrane of mozambique tilapia (Oreochromis mossambicus) embryos and larvae during seawater adaptation

1999 ◽  
Vol 202 (24) ◽  
pp. 3485-3495 ◽  
Author(s):  
J. Hiroi ◽  
T. Kaneko ◽  
M. Tanaka
Keyword(s):  
Fresh Water ◽  
Cell Morphology ◽  
Sea Water ◽  
Oreochromis Mossambicus ◽  
Yolk Sac ◽  
Chloride Cell ◽  
Chloride Cells ◽  
Seawater Adaptation ◽  
Accessory Cells

Changes in chloride cell morphology were examined in the yolk-sac membrane of Mozambique tilapia (Oreochromis mossambicus) embryos and larvae transferred from fresh water to sea water. By labelling chloride cells with DASPEI, a fluorescent probe specific for mitochondria, we observed in vivo sequential changes in individual chloride cells by confocal laser scanning microscopy. In embryos transferred from fresh water to sea water 3 days after fertilization, 75 % of chloride cells survived for 96 h, and cells showed a remarkable increase in size. In contrast, the cell size did not change in embryos and larvae kept in fresh water. The same rate of chloride cell turnover was observed in both fresh water and sea water. Using differential interference contrast (DIC) optics and whole-mount immunocytochemistry with anti-Na(+)/K(+)-ATPase, we classified chloride cells into three developmental stages: a single chloride cell without an apical pit, a single chloride cell with an apical pit, and a multicellular complex of chloride and accessory cells with an apical pit. DIC and immunofluorescence microscopy revealed that single chloride cells enlarged and were frequently indented by newly differentiated accessory cells to form multicellular complexes during seawater adaptation. These results indicate that freshwater-type single chloride cells are transformed into seawater-type multicellular complexes during seawater adaptation, suggesting plasticity in the ion-transporting functions of chloride cells in the yolk-sac membrane of tilapia embryos and larvae.

Transport properties of cultured branchial epithelia from freshwater rainbow trout: a novel preparation with mitochondria-rich cells

2000 ◽  
Vol 203 (10) ◽  
pp. 1523-1537 ◽  
Author(s):  
M. Fletcher ◽  
S.P. Kelly ◽  
P. Part ◽  
M.J. O'Donnell ◽  
C.M. Wood
Keyword(s):  
Rainbow Trout ◽  
Flux Ratio ◽  
Culture Conditions ◽  
Chloride Cells ◽  
Pavement Cells ◽  
Ratio Criterion ◽  
Peg 4000 ◽  
Tight Epithelia ◽  
Gill Filaments

A new double-seeded insert (DSI) technique is described for culture of branchial epithelial preparations from freshwater rainbow trout on filter supports. DSI epithelia contain both pavement cells and mitochondria-rich (MR) cells (15.7+/−2.5 % of total cell numbers). MR cells occur singly or in clusters, are voluminous, open apically to the ‘external environment’ and exhibit ultrastructural characteristics similar to those found in the ‘chloride cells’ of freshwater fish gills. After 6–9 days in culture with Leibovitz's L-15 medium on both surfaces (symmetrical conditions), transepithelial resistance (TER) stabilized at values as high as 34 k capomega cm(2), indicative of electrically ‘tight’ epithelia. The density of MR cells, the surface area of their clusters and transepithelial potential (TEP; up to +8 mV basolateral positive, mean +1.9+/−0.2 mV) were all positively correlated with TER. In contrast, preparations cultured using an earlier single-seeded insert (SSI) technique contained only pavement cells and exhibited a negligible TEP under symmetrical conditions. Na(+)/K(+)-ATPase activities of DSI preparations were comparable with those in gill filaments, but did not differ from those of SSI epithelia. Replacement of the apical medium with fresh water to mimic the in vivo situation (asymmetrical conditions) induced a negative TEP (−6 to −15 mV) and increased permeability to the paracellular marker PEG-4000. Under symmetrical conditions, unidirectional Na(+) and Cl(−) fluxes were in balance, and there was no active transport by the Ussing flux ratio criterion. Under asymmetrical conditions, there were large effluxes, small influxes and evidence for active Cl(−) uptake and Na(+) extrusion. Unidirectional Ca(2+) fluxes were only 0.5-1.0 % of Na(+) and Cl(−) fluxes; active net Ca(2+) uptake occurred under symmetrical conditions and active net extrusion under asymmetrical conditions. Thus, DSI epithelia exhibit some of the features of the intact gill, but improvements in culture conditions are needed before the MR cells will function as true freshwater ‘chloride cells’.

Characterization of branchial transepithelial calcium fluxes in freshwater trout, Salmo gairdneri

1988 ◽  
Vol 254 (3) ◽  
pp. R491-R498 ◽  
Author(s):  
S. F. Perry ◽  
G. Flik
Keyword(s):  
Active Transport ◽  
Calcium Uptake ◽  
Chloride Cells ◽  
Salmo Gairdneri ◽  
Apical Surface ◽  
Electrochemical Gradient ◽  
Plasma Membrane Vesicles ◽  
Pavement Cells ◽  
Calcium Fluxes

Experiments were performed to determine whether gill transepithelial calcium fluxes in the freshwater trout (Salmo gairdneri) are passive or require active transport and to characterize the mechanisms involved. A comparison of the in vivo unidirectional flux ratios with the flux ratios calculated according to the transepithelial electrochemical gradients revealed that calcium uptake from the water requires active transport of Ca2+. The inhibition of calcium uptake by external lanthanum, the specific deposition of lanthanum on the apical surface of chloride cells, and the favorable electrochemical gradient for calcium across the apical membrane suggest that the initial step in branchial calcium uptake is the passive entry of calcium into the cytosol of chloride cells through apical channels that are permeable to calcium. The study of gill basolateral plasma membrane vesicles demonstrated the existence of a high-affinity calmodulin-dependent calcium-transporting system [half-maximal Ca2+ concentration (K0.5) = 160 nM, Vmax = 1.86 nmol.min-1.mg protein-1]. This system actively transports calcium from the cytosol of chloride cells into the plasma against a sizeable electrochemical gradient, thereby completing the transepithelial uptake of calcium. Calcium efflux occurs passively through paracellular pathways between chloride cells and adjacent pavement cells or between neighboring pavement cells.

scholarly journals Lophiosilurus alexandri, a sedentary bottom fish, adjusts its physiological parameters to survive to hypoxia condition.

2021 ◽  
Author(s):  
Livia de Assis Porto ◽  
Rafael Magno Costa Melo ◽  
Suzane Lilian Beier ◽  
Ronald Kennedy Luz ◽  
Gisele Cristina Favero
Keyword(s):  
Chloride Cells ◽  
Blood Capillary ◽  
Blood Gas ◽  
Hypoxia Exposure ◽  
Physiological Variables ◽  
Significant Difference ◽  
Undifferentiated Cells ◽  
Morphology And Morphometry ◽  
Hematological And Biochemical Parameters ◽  
Mucus Cells

Abstract We investigated blood gas, hematological and biochemical parameters and gill morphology and morphometry of Lophiosilurus alexandri juveniles submitted to hypoxia for 48 hours followed by recovery for 48 hours. A total of 48 juveniles (360.0 ± 141.6 g) were distributed among eight tanks (120 L) and subjected to hypoxia condition (water with dissolved oxygen at 2.12 ± 0.90 mg L− 1) or normoxia (at 5.60 ± 0.31 mg L− 1). Blood gas values (pH, PvCO2, PvO2, sO2, HCO3−, stHCO3− and base excess) in hypoxia were significantly different from normoxia, while lactate and the electrolytes (K+, Na+, Cl−, Ca2+ and HCO3−) there was no significant change among treatments. The erythrocytes differed significantly between hypoxia and normoxia at 24 h of recovery, while for hemoglobin and hematocrit there were no significant differences. There was a significant difference in glucose, triglycerides, and cholesterol for both normoxia and hypoxia, while plasma protein remained unchanged. All gill components (epithelial cells, erythrocytes, pillar cells, mucus cells, chloride cells, undifferentiated cells, and blood capillary lumen) differed significantly between hypoxia and normoxia. A reduction in the length of the primary lamella was observed in the hypoxia and recovery treatments, when compared to normoxia. The secondary branchial lamella showed no significant difference for both treatments. In general, juveniles of L. alexandri adapted well to hypoxia exposure for 48 h, as they were able to adjust most of their physiological variables to survive this stress condition and return to normoxia within 48 h.

Ultrastructure and distribution dynamics of chloride cells in tilapia larvae in fresh water and sea water

1999 ◽  
Vol 297 (1) ◽  
pp. 119-130 ◽  
Author(s):  
A. J. H. van der Heijden ◽  
J. C. A. van der Meij ◽  
G. Flik ◽  
S. E. Wendelaar Bonga
Keyword(s):  
Fresh Water ◽  
Sea Water ◽  
Chloride Cells ◽  
Distribution Dynamics
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