Duplicated CFTR isoforms in eels diverged in regulatory structures and osmoregulatory functions.
Two cystic fibrosis transmembrane conductance regulator (CFTR) isoforms, CFTRa and CFTRb, were cloned in Japanese eel and their structures and functions were studied in different osmoregulatory tissues in freshwater (FW) and seawater (SW) eels. Molecular phylogenetic results suggested that the CFTR duplication in eels occurred independently of the duplication event in salmonid. CFTRa was expressed in the intestine and kidney and downregulated in both tissues in SW eels, while CFTRb was specifically expressed in the gill and greatly upregulated in SW eels. Structurally, the CFTR isoforms are similar in most functional domains except the regulatory R domain, where the R domain of CFTRa is similar to that of human CFTR but the R domain of CFTRb is unique in having high intrinsic negative charges and fewer phosphorylation sites, suggesting divergence of isoforms in terms of gating properties and hormonal regulation. Immunohistochemical results showed that CFTR was localized on the apical regions of SW ionocytes, suggesting a Cl(-) secretory role as in other teleosts. In intestine and kidney, however, immunoreactive CFTR was mostly found in the cytosolic vesicles in FW eels, indicating that Cl(-) channel activity could be low at basal conditions, but could be rapidly increased by membrane insertion of the stored channels. Guanylin (GN), a known hormone that increases CFTR activity in mammalian intestine, failed to redistribute CFTR and to affect its expression in eel intestine. The results suggested that GN-independent CFTR regulation is present in eel intestine and kidney.
Atmosphere and Ocean Research Institute, The University of Tokyo, Chiba, Japan. Electronic address: martywong@aori.u-tokyo.ac.jp.
Marty Kwok-Shing
MK
Wong
Atmosphere and Ocean Research Institute, The University of Tokyo, Chiba, Japan.
Supriya
S
Pipil
Department of Biological Sciences, Tokyo Institute of Technology, Yokohama, Japan.
Akira
A
Kato
Atmosphere and Ocean Research Institute, The University of Tokyo, Chiba, Japan.
Yoshio
Y
Takei
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D004524:Eels / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D005618:Fresh Water; D020869:Gene Expression Profiling / Q000379:methods; D020131:Genes, Duplicate / Q000235:genetics; D014644:Genetic Variation; D005880:Gills / Q000378:metabolism; D007150:Immunohistochemistry; D007413:Intestinal Mucosa / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D064587:Osmoregulation / Q000235:genetics* / Q000502:physiology; D010766:Phosphorylation; D010802:Phylogeny; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012623:Seawater; D017386:Sequence Homology, Amino Acid
D029941:Fish Proteins; D020033:Protein Isoforms; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
CFTR; Chloride secretion; Gene duplication; Guanylin; Isoform; Osmoregulation