Peanut lectin binds to a subpopulation of mitochondria-rich cells in the rainbow trout gill epithelium

Abstract

Fluorescently labeled peanut lectin agglutinin (PNA-FITC) was used to identify a subtype of mitochondria-rich (MR) cells in the gills of freshwater rainbow trout. In situ binding of PNA-FITC was visualized by inverted fluorescence microscopy and found to bind to cells on the trailing edge of the filament epithelium as demonstrated by differential interference contrast optics. The amount of PNA-FITC binding on the filament epithelium increased with cortisol pretreatment concomitant with an increased chloride cell fractional area as demonstrated by scanning electron microscopy. Dispersed gill cells were isolated by trypsinization and separated using a discontinuous Percoll density gradient. Cells migrating to the 1.06-1.09 g/ml interface were found to be MR as demonstrated by staining with the vital mitochondrial dye 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide and transmission electron microscopy (TEM). However, only approximately 40% of the MR cells were found to bind PNA-FITC. Cortisol pretreatment increased the relative numbers of MR cells isolated from the dispersed gill cell population, but the relative proportions of PNA binding cells remained unchanged. Ultrastructural analysis of isolated cells in the TEM demonstrated that the MR cell fraction was comprised of a mixed population of chloride cells and pavement cells.

Isolation and fractionation of gill cells from freshwater (Lasmigona costata) and seawater (Mesodesma mactroides) bivalves for use in toxicological studies with copper

“…The methodologies employed for isolation and fractionation of cells from the gills of the freshwater bivalve L. costata and the seawater clam M. mactroides were found to be effective, as previously reported for fish (Goss et al 2001;Galvez et al 2002;Tse et al 2006). Also, the application of two different approaches (mitochondria density and Na ?…”

“…functional characterization of cell membrane transporters related to Na ? uptake (Goss et al 2001;Galvez et al 2002;Reid et al 2003), and particularly in analyzing the ionoregulatory disrupting effects of metals such as Cu (Goss et al 2011). Cu is now generally recognized to be an ionoregulatory toxicant in aquatic invertebrates (Pinho et al 2007;Lopes et al 2011a, b) as well as in fish.…”

The involvement of SLC26 anion transporters in chloride uptake in zebrafish (Danio rerio) larvae

“…In rainbow trout (Goss and Perry, 1993;Perry and Goss, 1994), brown bullhead (Ictalurus nebulosus) (Goss et al, 1992; and tilapia (Oreochromis mossambicus) (Chang et al, 2003), the rates of Cl -uptake (and in some cases base excretion) are significantly correlated with the numbers of MR cells in contact with the water. In light of the finding of specific subtypes of MR cells (Pisam et al, 1987;Chang et al, 2001;Goss et al, 2001;Hiroi et al, 2005;Laurent et al, 2006), models have been developed in which specific functions are assigned to the various MR cell types. By analogy with the β-intercalated cell of the mammalian renal collecting duct, one of the branchial MR cell subtypes is thought to function as a base-secreting cell with an apical membrane Cl ) or (B) the effects of a 24 h incubation of 7 d.p.f.…”

Na+/K+-ATPase α-isoform switching in gills of rainbow trout (Oncorhynchus mykiss) during salinity transfer

“…Trout gills are composed of multiple cell types including respiratory pavement cells and ion-regulating mitochondriarich (MR; chloride) cells. Recently, two distinct populations of MR cells have been isolated from trout gills (Galvez et al, 2002;Goss et al, 2001) and there is accumulating evidence that these two MR cell populations may differ in their complement of ion transporters; thus it seems likely that these two cell populations may be differentially proliferated during freshwater and seawater acclimation. Differential changes in cell proliferation during salinity transfer may explain some of the observed changes in Na + /K + -ATPase α-isoform expression (Fig.·7A,B).…”