TY - JOUR
T1 - Defects in Cholangiocyte Fibrocystin Expression and Ciliary Structure in the PCK Rat
AU - Masyuk, Tatyana V.
AU - Huang, Bing Q.
AU - Ward, Christopher J.
AU - Masyuk, Anatoliy I.
AU - Yuan, David
AU - Splinter, Patrick L.
AU - Punyashthiti, Rachaneekorn
AU - Ritman, Eric L.
AU - Torres, Vicente E.
AU - Harris, Peter C.
AU - Larusso, Nicholas F.
PY - 2003/11
Y1 - 2003/11
N2 - 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 Pkhdl 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 Pkhdl 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 Pkhdl by a germ line mutation in the PCK rat or by siRNA in IBDUs results in abnormalities in ciliary morphology and possibly biliary cystogenesis.
AB - 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 Pkhdl 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 Pkhdl 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 Pkhdl 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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U2 - 10.1016/j.gastro.2003.09.001
DO - 10.1016/j.gastro.2003.09.001
M3 - Article
C2 - 14598246
AN - SCOPUS:0142168748
SN - 0016-5085
VL - 125
SP - 1303
EP - 1310
JO - Gastroenterology
JF - Gastroenterology
IS - 5
ER -