Sodium/proton antiporter in the euryhaline crab Carcinus maenas: molecular cloning, expression and tissue distribution.
Gill epithelial cells of euryhaline crustaceans demonstrate net inward transport of sodium ions, possibly via apical Na+/H+ antiporters, Na+/K+/2Cl- cotransporters or Na+ channels working in series with the basolateral Na(+) + K(+)-ATPase. We have identified and sequenced the cDNA coding for a crustacean Na+/H+ antiporter, starting with mRNA isolated from gills of the euryhaline green shore crab Carcinus maenas. The complete 2595-base-pair cDNA includes an open reading frame coding for a 673-amino-acid protein. A search of GenBank revealed more than 20 high-scoring matches, all Na+/H+ antiporter sequences from mammalian, amphibian, teleost and nematode species. Injection of Xenopus laevis oocytes with cRNA transcribed from the cloned crab sequence substantially enhanced Na(+)-dependent H+ efflux from the oocytes. Analysis of crab tissue antiporter mRNA levels by semi-quantitative reverse transcription-polymerase chain reaction revealed that posterior and anterior gills of Carcinus maenas expressed this antiporter the most strongly, followed in decreasing order by skeletal muscle, hepatopancreas, hypodermis and heart. Hydropathy and transmembrane alpha-helix analysis suggested a 10-helix membrane-spanning topology of the antiporter protein. It is clear from this study that Carcinus maenas gills vigorously transcribe a gene coding for a Na+/H+ antiporter. Whether these gills also express a gene coding for an epithelial Na+ channel or Na+/K+/2Cl- cotransporter remains to be demonstrated.
Towle
D W
DW
Department of Biology, Lake Forest College, IL 60045, USA. towle@lfc.edu
Rushton
M E
ME
Heidysch
D
D
Magnani
J J
JJ
Rose
M J
MJ
Amstutz
A
A
Jordan
M K
MK
Shearer
D W
DW
Wu
W S
WS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003386:Brachyura; D018076:DNA, Complementary / Q000032:analysis / Q000737:chemistry; D015870:Gene Expression; D005880:Gills / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000302:isolation & purification; D016415:Sequence Alignment; D017421:Sequence Analysis; D017923:Sodium-Hydrogen Exchangers / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D017923:Sodium-Hydrogen Exchangers