Tight junctions are sensitive to peptides eliminated in the urine.
We prepare an extract of dog urine (DLU) that, when applied to monolayers of MDCK cells (epithelial, derived from a normal dog), enhances the transepithelial electrical resistance (TER) in a dose-dependent manner. This increase is not reflected in variations of the linear amount of TJ nor in changes of the pattern of junctional strands as observed in freeze fracture replicas, nor in the distribution of claudin 1 (a membrane protein of the TJ) nor ZO-1 (a TJ-associated protein). A preliminary characterization of the active component of DLU indicates that it weighs 30-50 kDa, bears a net negative electric charge, and is destroyed by type I protease but not by 10-min boiling. DLUs prepared from human, dog, rabbit and cat are effective on MDCK cells. However, dog DLU increases TER in MDCK (dog) as well as LLCPK1 (pig) monolayers, but not in other epithelial cell lines such as LLCRK1 (rabbit), PTK2 (kangaroo) and MA-104 (monkey), nor in the endothelial cell line CPA47 (cow). Given that in its transit from the glomerulus to the urinary bladder the filtrate increases its concentration by more than two orders of magnitude, the substance(s) we report may act at increasingly higher concentrations in each segment, and afford a potential clue to the progressive increase of TER across the walls of the nephron from the proximal to the collecting duct.
Gallardo
J M
JM
Orient Center of Research, Mexican Institute of Social Security, Puebla, Puebla, México.
Hernández
J M
JM
Contreras
R G
RG
Flores-Maldonado
C
C
González-Mariscal
L
L
Cereijido
M
M
D000818:Animals; D002415:Cats; D002417:Cattle; D002478:Cells, Cultured; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D004848:Epithelium / Q000187:drug effects / Q000502:physiology / Q000648:ultrastructure; D000882:Haplorhini; D006358:Hot Temperature; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D007614:Macropodidae; D008297:Male; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology / Q000652:urine*; D011817:Rabbits; D012016:Reference Values; D012680:Sensitivity and Specificity; D013552:Swine; D019108:Tight Junctions / Q000187:drug effects / Q000502:physiology*; D014556:Urine / Q000737:chemistry*
D010455:Peptides