title: Ion-motive ATPases and active, transbranchial NaCl uptake in the red freshwater crab, Dilocarcinus pagei (Decapoda, Trichodactylidae).
abstract: The present investigation examined the microanatomy and mRNA expression and activity of ion-motive ATPases, in anterior and posterior gills of a South American, true freshwater crab, Dilocarcinus pagei. Like diadromous crabs, the anterior gills of this hololimnetic trichodactylid exhibit a highly attenuated (2-5 microm), symmetrical epithelium on both lamellar surfaces. In sharp contrast, the posterior gill lamellar epithelia are markedly asymmetrical. Their proximal side consists of thick (18-20 microm) cells, displaying features typical of a transporting epithelium, while the distal epithelium is thin (3-10 microm) and formed entirely by apical pillar cell flanges. Both anterior and posterior gills express Na+/K+- and V-ATPases. Phylogenetic analysis of partial cDNA sequences for the Na+/K+-ATPase alpha-subunit and V-ATPase B-subunit among various crab species confirmed the previous classification and grouping of D. pagei based on morphological criteria. Semi-quantitative RT-PCR clearly showed that mRNA for both ion pump subunits is more intensely expressed in posterior gills. Na+/K+-ATPase activity in the posterior gills was nearly fourfold that of anterior gills, while V-ATPase and F-ATPase activities did not differ. A negative short-circuit current (Isc) was measured using the distal side of split, posterior gill lamellae, mounted in a modified Ussing chamber and perfused symmetrically with identical hemolymph-like salines. Although hemolymph-side ouabain did not affect this current, concanamycin significantly reduced Isc without altering preparation conductance, suggesting V-ATPase-driven Cl- absorption on the distal side of the posterior gill lamellae, as known to occur in diadromous crabs adapted to freshwater. These findings suggest that active Na+ uptake predominates across the thick proximal epithelium, and Cl- uptake across the thin, distal epithelium of the posterior gill lamellae.
authors.affiliation: Department of Animal Physiology, University of Osnabrueck, 49076 Osnabrueck, Germany. weihrauch@biologie.uni-osnabrueck.de
authors.forename: Dirk
authors.initials: D
authors.lastname: Weihrauch
authors.forename: John Campbell
authors.initials: JC
authors.lastname: McNamara
authors.forename: David W
authors.initials: DW
authors.lastname: Towle
authors.forename: Horst
authors.initials: H
authors.lastname: Onken
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001693:Biological Transport, Active / Q000502:physiology; D003386:Brachyura / Q000033:anatomy & histology / Q000378:metabolism*; D001938:Brazil; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004594:Electrophysiology; D004848:Epithelium / Q000378:metabolism; D005618:Fresh Water; D005880:Gills / Q000033:anatomy & histology* / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D010042:Ouabain; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012965:Sodium Chloride / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D012965:Sodium Chloride; D010042:Ouabain; D025262:Vacuolar Proton-Translocating ATPases; D000254:Sodium-Potassium-Exchanging ATPase