The effect of highly alkaline water (pH 9.5) on the morphology and morphometry of chloride cells and pavement cells in the gills of the freshwater rainbow trout: relationship to ionic transport and ammonia excretion

2000 ◽  
Vol 78 (2) ◽  
pp. 307-319 ◽  
Author(s):  
Pierre Laurent ◽  
Michael P Wilkie ◽  
Claudine Chevalier ◽  
Chris M Wood
Keyword(s):  
Rainbow Trout ◽  
Surface Area ◽  
Ammonia Excretion ◽  
Chloride Cells ◽  
Apical Surface ◽  
Influx Rate ◽  
Pavement Cells ◽  
Alkaline Water ◽  
Partial Restoration ◽  
Water Ph

Exposure of rainbow trout (Oncorhynchus mykiss) to alkaline water (pH 9.5) impairs ammonia excretion (JAmm) and gill-mediated ion-exchange processes, as characterized by decreased Cl- (JC1in) and Na+ influx (JNain) across the gill. Scanning electron microscopy suggested that the depression of JC1in was concomitant with an early decrease in the population of the most active chloride cells (CCs), partly compensated for by an increasing number of immature CCs. However, within 72 h after the onset of exposure to alkaline water, there was a 2-fold increase in the fractional apical surface area of CCs that paralleled complete recovery of the maximal Cl- influx rate (JC1max). These results suggest that recovery of JC1max was associated with greater CC surface area, resulting in more transport sites on the gill epithelium. Morphometric analysis of the outermost layer of pavement cells on the lamellar epithelium showed a greater density of microvilli during exposure to alkaline water, which may have contributed to partial restoration of the number of Na+ transport sites (JNamax). Finally, the blood-to-water gill-diffusion distance decreased by 27% after 72 h at pH 9.5, and likely contributed to progressive restoration of ammonia excretion in alkaline water.

Gill morphometry and fish osmoregulation

1989 ◽  
Vol 67 (12) ◽  
pp. 3055-3063 ◽  
Author(s):  
Pierre Laurent ◽  
Nadra Hebibi
Keyword(s):  
Rainbow Trout ◽  
Surface Area ◽  
Surface Density ◽  
Tap Water ◽  
Chloride Cells ◽  
Apical Surface ◽  
Ionic Regulation ◽  
Epithelial Surface ◽  
Rainbow Trout Oncorhynchus Mykiss ◽  
Gill Lamellae

Morphofunctional parameters were studied during adaptation of rainbow trout (Oncorhynchus mykiss) to different ionic environments: Strasbourg tap water, ion-poor water, and artificial seawater. The gill lamellae displayed large changes in size. Surface area of individual lamellae increased in trout acclimated to ion-poor water or seawater. Conversely, the harmonic mean thickness of the lamellar epithelium decreased in seawater, and to an even greater extent in ion-poor water. The apical surface area of individual branchial filament chloride cells, the number of these cells, and their apical surface density per unit of filament epithelial surface area were calculated in these three conditions. These variables did not differ significantly in Strasbourg tap water or seawater, but increased greatly in ion-poor water. These results are discussed in relation to gill permeability and ionic regulation in fish.

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 Adaptational Responses of Rainbow Trout to Lowered External Nacl Concentration: Contribution of the Branchial Chloride Cell

1989 ◽  
Vol 147 (1) ◽  
pp. 147-168 ◽  
Author(s):  
STEVE F. PERRY ◽  
PIERRE LAURENT
Keyword(s):  
Rainbow Trout ◽  
Surface Area ◽  
Fresh Water ◽  
Primary Response ◽  
Chloride Cell ◽  
Whole Body ◽  
Salmo Gairdneri ◽  
Apical Surface ◽  
Ionic Fluxes ◽  
Area 5

1. Whole-body ionic fluxes and gill chloride cell (CC) morphology were monitored in rainbow trout (Salmo gairdneri) exposed acutely or chronically to natural fresh water (NFW; [Na+]=0.120 mmoll−1; [Cr]=0.164 mmoll−1) or artificially prepared fresh water with reduced [NaCl] (AFW; [Na+]=0.017 mmoll−1; [CT]=0.014 mmoll−1). 2. Net fluxes of Na+ (JnetNa) and Cl− (JnetCl) became extremely negative (indicating net NaCl loss to the environment) upon immediate exposure to AFW exclusively as a result of reduced NaCl influx (JinNa and JinNa). JnetNa and JnetCl were gradually restored to control rates during prolonged (30 days) exposure to AFW. 3. The restoration of JnetCl in AFW was due both to increased JinCl and to reduced Cl− efflux (JoutCl) whereas the primary response contributing to the restoration of JnetNa a t was an increase of JNain. 4. The total apical surface area of branchial CCs exposed to the external environment increased markedly after 24 h in AFW and remained elevated for 1 month as a consequence of enlargement of individual CCs and, to a lesser extent, increased CC density. JinNa and JinNa were correlated significantly with total CC apical surface area. 5. Plasma cortisol levels rose transiently in fish exposed to AFW. Treatment of NFW-adapted fish with cortisol for 10 days (a protocol known to cause CC proliferation) caused pronounced increases in JinCl and JinNa, as measured in both NFW and AFW. 6. These results suggest that an important adaptational response of rainbow trout to low environmental [NaCl] is cortisol-mediated enlargement of branchial epithelial CCs which, in turn, enhances the NaCl-transporting capacity of the gill as a result of the proliferation of Na+ and Cl− transport sites.

Ammonia excretion in rainbow trout (Oncorhynchus mykiss): evidence for Rh glycoprotein and H+-ATPase involvement

2007 ◽  
Vol 31 (3) ◽  
pp. 463-474 ◽  
Author(s):  
C. Michele Nawata ◽  
Carrie C. Y. Hung ◽  
Tommy K. N. Tsui ◽  
Jonathan M. Wilson ◽  
Patricia A. Wright ◽  
...  
Keyword(s):  
Boundary Layer ◽  
Rainbow Trout ◽  
Ammonia Excretion ◽  
Pavement Cell ◽  
New Members ◽  
Pavement Cells ◽  
Cdna Sequences ◽  
Rainbow Trout Oncorhynchus Mykiss ◽  
Ammonia Transport ◽  
Genes Encoding

Branchial ammonia transport in freshwater teleosts is not well understood. Most studies conclude that NH3 diffuses out of the gill and becomes protonated to NH4+ in an acidified gill boundary layer. Rhesus (Rh) proteins are new members of the ammonia transporter superfamily and rainbow trout possess genes encoding for Rh30-like1 and Rhcg2. We identified seven additional full-length trout Rh cDNA sequences: one Rhag and two each of Rhbg, Rhcg1, and Rh30-like. The mRNA expression of Rhbg, Rhcg1, and Rhcg2 was examined in trout tissues (blood, brain, eye, gill, heart, intestine, kidney, liver, muscle, skin, spleen) exposed to high external ammonia (HEA; 1.5 mmol/l NH4HCO3, pH 7.95, 15°C). Rhbg was expressed in all tissues, Rhcg1 was expressed in brain, gill, liver, and skin, and Rhcg2 was expressed in gill and skin. Brain Rhbg and Rhcg1 were downregulated, blood Rh30-like and Rhag were downregulated, and skin Rhbg and Rhcg2 were upregulated with HEA. After an initial uptake of ammonia into the fish during HEA, excretion was reestablished, coinciding with upregulations of gill Rh mRNA in the pavement cell fraction: Rhcg2 at 12 and 48 h, and Rhbg at 48 h. NHE2 expression remained unchanged, but upregulated H+-ATPase (V-type, B-subunit) and downregulated carbonic anhydrase (CA2) expression and activity were noted in the gill and again expression changes occurred in pavement cells, and not in mitochondria-rich cells. Together, these results indicate Rh glycoprotein involvement in ammonia transport and excretion in the rainbow trout while underscoring the significance of gill boundary layer acidification by H+-ATPase.

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.

Changes in the apical surface of chloride cells following acclimation of lampreys to seawater

1996 ◽  
Vol 270 (1) ◽  
pp. R125-R133 ◽  
Author(s):  
H. Bartels ◽  
A. Moldenhauer ◽  
I. C. Potter
Keyword(s):  
Electron Microscopy ◽  
Paracellular Permeability ◽  
Passive Movement ◽  
Chloride Cells ◽  
Apical Surface ◽  
Full Width ◽  
Gill Epithelium ◽  
Pavement Cells ◽  
Paracellular Shunt ◽  
Scanning Electron

Scanning electron microscopy (SEM) was used to study the changes that occur in the morphological relationships between chloride and pavement cells in the gills during acclimation of young adult lampreys to seawater. Because chloride cells are located predominantly between lamellae and are thus obscured from view, the lamellae were removed with the use of a micromanipulator installed in a SEM. In gills of animals maintained in river water, chloride cells could then be seen to be dislike and typically to form single rows between successive lamellae. After acclimation to seawater, the apical surfaces of chloride cells lose their microvilli and change in shape from small circles to rectangles that extend the full width between successive lamellae. These changes result in an increase in the length of the paracellular pathway between chloride cells. Previous work has shown that the number of strands of the zonulae occludentes sealing this pathway declines under these conditions. This presumably leads to an increase in paracellular permeability of the gill epithelium, thereby providing the low-resistance paracellular shunt required for the passive movement of sodium into the environment during osmoregulation in seawater. The above changes are reversed by transfer of lampreys downward to 10% seawater.

Isolation and characterization of mitochondria-rich cell types from the gill of freshwater rainbow trout

2002 ◽  
Vol 282 (3) ◽  
pp. R658-R668 ◽  
Author(s):  
Fernando Galvez ◽  
Scott D. Reid ◽  
Guy Hawkings ◽  
Greg G. Goss
Keyword(s):  
Rainbow Trout ◽  
Iron Particle ◽  
Cell Types ◽  
Chloride Cells ◽  
Transmission Electron Microscopy Analysis ◽  
Pavement Cells ◽  
Isolation And Characterization ◽  
Transmission Electron ◽  
Electron Microscopy Analysis ◽  
Magnetic Cell Separation

A magnetic cell separation technique (MACS) was developed for isolating and characterizing peanut lectin agglutinin positive (PNA+) cells from rainbow trout gills. Percoll density separated mitochondria-rich (MR) cells were serially labeled with PNA-FITC and an anti-FITC antibody covalently coupled to a 50-nm iron particle and then applied to a magnetic column. PNA+ MR cells were enriched to >95% purity. Transmission electron microscopy analysis of both the PNA+ and PNA negative (PNA−) fraction showed that PNA binds to MR chloride cells while the PNA− cell fraction is comprised of MR cells with features characteristic of pavement cells. Western blotting demonstrated that both PNA+ and PNA− fractions had high levels of Na+-K+-ATPase and Sco1 expression; however, relative expression of H+-ATPase in PNA+ and PNA− cells demonstrated that untreated fish had twofold higher H+-ATPase levels in PNA− cells relative to the PNA+ cells. Furthermore, hypercapnic acidosis significantly increased the relative H+-ATPase expression on PNA− cells only, whereas metabolic alkalosis had no significant effect.

scholarly journals Histology and Surface Ultra-Structure of the Gill of A Minor Carp, Labeo bata (Hamilton)

2017 ◽  
Vol 9 (2) ◽  
pp. 201-208
Author(s):  
I. Samajdar ◽  
D. K. Mandal
Keyword(s):  
Light And Electron Microscopy ◽  
Chloride Cells ◽  
Apical Surface ◽  
Mucous Cells ◽  
Pavement Cells ◽  
Stratified Epithelium ◽  
Labeo Bata ◽  
Secondary Lamellae ◽  
Secondary Lamella ◽  
A Minor

The histology and surface ultra-structures of the gill epithelium of Labeo bata has been examined through light and electron microscopy. Gill arches bear double rows of primary lamellae and each primary lamella is provided with two rows of delicate rectangular secondary lamellae on its upper and lower surfaces. Gill arches and filaments are lined by thick stratified epithelium while the epithelium of secondary lamella is made up of thin single layered pavement cells. The thin epithelium is supported and kept apart from each other by the pillar cells. The pavement cells are polygonal with well-defined cell boundary and microridges on its apical surface. The epithelial cells are interspersed by large chloride cells located at the base of secondary lamellae and PAS positive mucous cells. Mucous cells are found abundant at the base and tip of the primary lamellae.  Taste buds are found on the concave anterior faces of gill arches. The short and soft gill rakers are covered with stratified epithelium. This finding demonstrated that the fish has well developed respiratory system matching to its active life.

Sign in / Sign up

Export Citation Format

Share Document