Two splice variants of claudin-10 in the kidney create paracellular pores with different ion selectivities.
Members of the large claudin family of tight junction (TJ) proteins create the differences in paracellular conductance and charge selectivity observed among different epithelia. Previous studies demonstrated that ionic charge selectivity is influenced by acidic or basic amino acids on the first extracellular domain of claudins. We noted two alternatively spliced variants of claudin-10 in the database, 10a and 10b, which are predicted to encode two different first extracellular domains and asked whether this might be a novel mechanism to generate two different permselectivities from a single gene. Using quantitative PCR, we found that claudin-10b is widely expressed among tissues including the kidney; however, claudin-10a is unique to the kidney. Using a nondiscriminating antibody, we found that claudin-10 (a plus b) is expressed in most segments of the nephron. In situ hybridization, however, showed that mRNA for 10a is concentrated in the cortex, and mRNA for 10b is more highly expressed in the medulla. Expression in Madin-Darby canine kidney (MDCK) II and LLC-PK1 cells reveals that both variants form low-resistance pores, and that claudin-10b is more selective for cations than claudin-10a. Charge-reversing mutations of cationic residues on 10a reveal positions that contribute to its anion selectivity. We conclude that alternative splicing of claudin-10 generates unique permselectivities and might contribute to the variable paracellular transport observed along the nephron.
Van Itallie
Christina M
CM
Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7545, USA. vitallie@med.unc.edu
Rogan
Sarah
S
Yu
Alan
A
Vidal
Lucia Seminario
LS
Holmes
Jennifer
J
Anderson
James M
JM
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D057167:Claudins; D004285:Dogs; D007477:Ions / Q000378:metabolism; D007672:Kidney Cortex / Q000502:physiology*; D007679:Kidney Medulla / Q000502:physiology*; D018374:LLC-PK1 Cells; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D013552:Swine; D019108:Tight Junctions / Q000378:metabolism
D057167:Claudins; D007477:Ions; D008565:Membrane Proteins; D020033:Protein Isoforms; C502009:claudin 10