Defects in cholangiocyte fibrocystin expression and ciliary structure in the PCK rat
TV Masyuk, BQ Huang, CJ Ward, AI Masyuk, D Yuan… - Gastroenterology, 2003 - Elsevier
TV Masyuk, BQ Huang, CJ Ward, AI Masyuk, D Yuan, PL Splinter, R Punyashthiti, EL Ritman…
Gastroenterology, 2003•ElsevierBACKGROUND & AIMS:: Recent studies have showed that proteins associated with
polycystic kidney disease (PKD) are expressed in cilia, linking this organelle and cyst
formation in the kidney, but involvement of cilia in PKD-related biliary cystogenesis has not
been shown. We investigated:(1) the expression of fibrocystin (a product of PKHD1, the
autosomal-recessive PKD [ARPKD] gene) in cholangiocyte cilia;(2) biliary cyst formation in
an orthologous rat model, PCK; and (3) the effect of Pkhd1 mutation on ciliary structure …
polycystic kidney disease (PKD) are expressed in cilia, linking this organelle and cyst
formation in the kidney, but involvement of cilia in PKD-related biliary cystogenesis has not
been shown. We investigated:(1) the expression of fibrocystin (a product of PKHD1, the
autosomal-recessive PKD [ARPKD] gene) in cholangiocyte cilia;(2) biliary cyst formation in
an orthologous rat model, PCK; and (3) the effect of Pkhd1 mutation on ciliary structure …
BACKGROUND & AIMS
Recent studies have showed that proteins associated with polycystic kidney disease (PKD) are expressed in cilia, linking this organelle and cyst formation in the kidney, but involvement of cilia in PKD-related biliary cystogenesis has not been shown. We investigated: (1) the expression of fibrocystin (a product of PKHD1, the autosomal-recessive PKD [ARPKD] gene) in cholangiocyte cilia; (2) biliary cyst formation in an orthologous rat model, PCK; and (3) the effect of Pkhd1 mutation on ciliary structure.
METHODS
Biliary cystogenesis was assessed by microcomputed tomography. Fibrocystin expression in cholangiocytes of isolated intrahepatic bile ducts (IBDUs) and liver cysts was analyzed by confocal and immunoelectron microscopy, and ciliary structure and length by scanning and transmission electron microscopy. Small interfering RNAs (siRNA) were used to examine the effect of fibrocystin loss on ciliary structure.
RESULTS
The biliary tree in the PCK rat was distorted markedly, showing multiple bile duct dilatation and focal budding. In normal IBDUs, each cholangiocyte had a single cilium that expressed fibrocystin. In contrast, cilia in the PCK rat were abnormal with bulbous extensions and diminished length, and were devoid of fibrocystin. In cholangiocytes of normal IBDUs, specific siRNA reduced Pkhd1 messenger RNA by 80%, the length of cilia by 41%, and fibrocystin ciliary expression to an undetectable level.
CONCLUSIONS
Our results indicate that fibrocystin is expressed in cholangiocyte cilia and that disruption of Pkhd1 by a germ line mutation in the PCK rat or by siRNA in IBDUs results in abnormalities in ciliary morphology and possibly biliary cystogenesis.
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