Mitochondria-rich cells in amphibian skin epithelium: Relationship of immuno- and peanut lectin labeling pattern and transport functions.

Mitochondria-rich cells are an integral component of the epidermis of amphibian skin and play a functional role. Whereas the principal cell compartment of the epithelium is specialized almost exclusively for active uptake of sodium, the mitochondria-rich cells perform other diverse ion-transport functions, including transport of Cl(-), H(+), HCO(3)(-) and organic molecules. These transporting functions differ in different species. Antibodies, such as those directed against band 3, H(+)-ATPase, and also peanut lectin (PNA), bind specifically to the mitochondria-rich cells, but do so differently in various species. Examination of these immunolocalizations and lectin labeling in the skin of over 10 amphibian species, including both Anurans and Urodeles, illustrate species-specific differences. The binding pattern and the transport capabilities of the skin in the various species do not show a universal correlation, they appear to be species specific and do not permit construction of a general scheme common to all the species studied. The mitochondria-rich cells of heterocellular epithelia and their roles in ion transport remain a subject that requires further studies to elucidate their particular functions within the framework of the whole epithelium.

Enzymatic changes in mitochondria-rich cells of Xenopus laevis skin epithelium are induced by ionic acclimation.

Morphological, biochemical and histochemical components of mitochondria-rich (MR) cells of skin epithelium of Xenopus laevis (Daudin) were investigated after acclimation in distilled water (DW) and mild solutions (50 mmol/l) of either NaCl or KCl for over 10 days. The animals readily acclimated to NaCl, but approximately 50% of the animals died in the KCl solution. Electrophysiological measurements confirmed the poor transport properties of skin in all conditions. Silver staining and exposure to methylene blue (MB) have shown that two types of MR cells can be distinguished, especially after KCl acclimation. Immunohistochemistry with the use of anti-band 3 polyclonal and anti H+-ATPase monoclonal antibodies demonstrated that band 3 and H+-ATPase enzymes were localized in MR cells in all conditions. H+-ATPase was greatly reduced during NaCl acclimation as verified with SDS gel electrophoresis. Intensity of the immunohistochemical staining differed between the various conditions of acclimation. Histochemical localization of carbonic anhydrase and alkaline phosphatase activities was more intense during NaCl acclimation. Morphological changes were also observed between the various acclimation conditions. The present findings substantiate the existence of at least two forms of MR cells in Xenopus skin epithelium but their functional significance remains to be established.

Skin epithelial transport and structural relationships in naturally metamorphosing Pelobates syriacus.

The onset of active Na(+) transport and activated Cl(-) conductance (G(Cl)) across the skin epithelium of Pelobates syriacus was investigated during natural ontogenetic development. Structural features, including band three and Peanut lectin bindings were tested in parallel and structure-function relationships were attempted. The 22 specimens studied were divided into two tadpole, three juvenile, and two adult stages, corresponding to the Taylor-Kollros standard table, in accordance with external morphology of their developmental stage. Onset of transepithelial electrical potential and drop in conductance occurred abruptly, coinciding with metamorphosis climax of tadpoles into juveniles at about stage XXI of development. Amiloride-sensitive Na(+) transport occurred a little later at stage XXIII, followed by the appearance of activated Cl(-) conductance, G(Cl). Parallel structural examination showed that skin MR cells occurred upon metamorphosis, as the tadpole integument transformed into the adult epithelium and could be associated with the occurrence of activated G(Cl). It was not related temporally with the appearance of band three protein in MR cells. Our findings support the association of G(Cl) with MR cells, whereas band three may only be a corollary of G(Cl) and not necessarily essential for its manifestation.

Structure-function relationships in the integument of Salamandra salamandra during ontogenetic development.

Morphological, cytological and transport properties of the integument of Salamandra salamandra were investigated during natural ontogenetic development, from birth to adult. Three stages were operationally defined: I, larvae, from birth to metamorphosis; II, metamorphosis (judged externally by the colour change and loss of the gills); and III, post-metamorphosis to adult. Pieces of skin were fixed at various stages for immunocytochemical examinations, and the electrical properties were investigated on parallel pieces. Distinct cellular changes take place in the skin during metamorphosis, and lectin (PNA, WGA and ConA) binding indicates profound changes in glycoprotein composition of cell membranes, following metamorphosis. Band 3 and carbonic anhydrase I (CA I) were confined to mitochondria-rich (MR)-like cells, and were detected only in the larval stage. CA II on the other hand, was detected both in MR-like and in MR cells following metamorphosis. The electrical studies show that the skin becomes more tight (transepithelial resistance increases) upon metamorphosis, followed by manifestation of amiloride-sensitive short-circuit current (I(SC)) indicating that functional Na+ uptake has been acquired. The skin of metamorphosed adults had no finite transepithelial Cl- conductance, and band 3 was not detected in its MR cells. The functional properties of MR-like and MR cells remain to be established.