title: Molecular characterization of a urea transporter in the gill of the gulf toadfish (Opsanus beta).
abstract: Urea excretion by the gulf toadfish (Opsanus beta) has been shown in previous studies to be a highly pulsatile facilitated transport, with excretion probably occurring at the gill. The present study reports the isolation of an 1800 base pair (kb) cDNA from toadfish gill with one open reading frame putatively encoding a 475-residue protein, the toadfish urea transporter (tUT). tUT, the first teleostean urea transporter cloned, has high homology with UTs (facilitated urea transporters) cloned from mammals, an amphibian and a shark, and most closely resembles the UT-A subfamily. When expressed in Xenopus laevis oocytes, tUT increased urea permeability (as measured by [(14)C]urea uptake) five- to sevenfold, and this permeability increase was abolished by phloretin, a common inhibitor of other UTs. Northern analysis using the 1.8 kb clone was performed to determine the tissue distribution and dynamics of tUT mRNA expression. Of six tissues examined (gill, liver, red blood cells, kidney, skin and intestine), only gill showed expression of tUT mRNA, with a predominant band at 1.8 kb and a minor band at 3.5 kb. During several points in the urea pulse cycle of toadfish (0, 4, 6, 12 and 18 h post-pulse), measured by excretion of [(14)C]urea into the water, gill mRNA samples were obtained. Expression of tUT mRNA was found to be largely invariant relative to expression of beta-actin mRNA over the pulse cycle. These results further confirm the gill localization of urea transport in the toadfish and suggest that tUT regulation (and the regulation of pulsatile urea excretion) is probably not at the level of mRNA control. The results are discussed in the context of the mechanisms of vasopressin-regulated UT-A in mammalian kidney and morphological data for the toadfish gill.
authors.affiliation: Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Science, NIEHS Marine and Freshwater Biomedical Sciences Center, University of Miami, Miami, FL 33149 USA. pwalsh@rsmas.miami.edu
authors.forename: P J
authors.initials: PJ
authors.lastname: Walsh
authors.forename: M J
authors.initials: MJ
authors.lastname: Heitz
authors.forename: C E
authors.initials: CE
authors.lastname: Campbell
authors.forename: G J
authors.initials: GJ
authors.lastname: Cooper
authors.forename: M
authors.initials: M
authors.lastname: Medina
authors.forename: Y S
authors.initials: YS
authors.lastname: Wang
authors.forename: G G
authors.initials: GG
authors.lastname: Goss
authors.forename: V
authors.initials: V
authors.lastname: Vincek
authors.forename: C M
authors.initials: CM
authors.lastname: Wood
authors.forename: C P
authors.initials: CP
authors.lastname: Smith
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D005399:Fishes / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D005880:Gills / Q000378:metabolism; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000235:genetics*; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014508:Urea / Q000378:metabolism*; D014982:Xenopus laevis; D000096982:Urea Transporters
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014508:Urea; D000096982:Urea Transporters