Salinity regulates claudin mRNA and protein expression in the teleost gill.
The teleost gill carries out NaCl uptake in freshwater (FW) and NaCl excretion in seawater (SW). This transformation with salinity requires close regulation of ion transporter capacity and epithelial permeability. This study investigates the regulation of tight-junctional claudins during salinity acclimation in fish. We identified claudin 3- and claudin 4-like immunoreactive proteins and examined their expression and that of select ion transporters by performing Western blot in tilapia (Oreochromis mossambicus) gill during FW and SW acclimation. Transfer of FW tilapia to SW increased plasma osmolality, which was corrected after 4 days, coinciding with increased gill Na+-K+-ATPase and Na+-K+-2Cl(-) cotransporter expression. Gill claudin 3- and claudin 4-like proteins were reduced with exposure to SW. Transfer to FW increased both claudin-like proteins. Immunohistochemistry shows that claudin 3-like protein was localized deep in the FW gill filament, whereas staining was found apically in SW gill. Claudin 4-like proteins are localized predominantly in the filament outer epithelial layer, and staining appears more intense in the gill of FW versus SW fish. In addition, tilapia claudin 28a and 30 genes were characterized, and mRNA expression was found to increase during FW acclimation. These studies are the first to detect putative claudin proteins in teleosts and show their localization and regulation with salinity in gill epithelium. The data indicate that claudins may be important in permeability changes associated with salinity acclimation and possibly the formation of deeper tight junctions in FW gill. This may reduce ion permeability, which is a critical facet of FW osmoregulation.
Tipsmark
Christian K
CK
Department of Zoology, North Carolina State University, Raleigh, North Carolina, USA. ckt@biology.sdu.dk
Baltzegar
David A
DA
Ozden
Ozkan
O
Grubb
Brenda J
BJ
Borski
Russell J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D001834:Body Water / Q000378:metabolism; D062506:Claudin-4; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D005618:Fresh Water; D005880:Gills / Q000378:metabolism*; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D009994:Osmolar Concentration; D012333:RNA, Messenger / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D054712:Salinity; D012623:Seawater; D028021:Sodium-Potassium-Chloride Symporters / Q000096:biosynthesis; D000254:Sodium-Potassium-Exchanging ATPase / Q000096:biosynthesis; D017210:Tilapia / Q000378:metabolism*
D062506:Claudin-4; D018076:DNA, Complementary; D008565:Membrane Proteins; D012333:RNA, Messenger; D028021:Sodium-Potassium-Chloride Symporters; D000254:Sodium-Potassium-Exchanging ATPase