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Doctoral Thesis
DOI
10.11606/T.5.2004.tde-04102005-111547
Document
Author
Full name
Luis Fernando Carvalho de Menezes
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2004
Supervisor
Committee
Onuchic, Luiz Fernando (President)
Aires, Margarida de Mello
Heilberg, Ita Pfeferman
Lopes, Anibal Gil
Reboucas, Nancy Amaral
Title in Portuguese
"Análise do padrão de expressão do produto de PKHD1, o gene mutado na doença renal policística autossômica recessiva"
Keywords in Portuguese
CÍLIOS/patologia
IMUNOHISTOQUÍMICA/métodos
ISOFORMAS DE PROTEÍNAS/análise
MICROSCOPIA IMUNOELETRÔNICA/métodos/ MICROSCOPIA DE FLUORESCÊNCIA/méto
PROTEÍNAS DE MEMBRANA/análise
RIM POLICÍSTICO AUTOSSÔMICO RECESSIVO/etiologia
RIM POLICÍSTICO AUTOSSÔMICO RECESSIVO/fisiopatologia
RIM POLICÍSTICO AUTOSSÔMICO RECESSIVO/genética
TÚBULOS COLETORES RENAIS/fisiopatologia
TÚBULOS COLETORES RENAIS/patologia
WESTERN BLOTTING/métodos
Abstract in Portuguese
O gene PKHD1, mutado na doença renal policística autossômica recessiva, apresenta um padrão de splicing complexo associado a múltiplos transcritos alternativos. Neste trabalho estudamos o perfil de expressão de seu produto, poliductina. Análises por western blot revelaram produtos putativos de membrana de > 440 kDa e ~230 kDa, e de ~140 kDa em frações solúveis de rim, fígado e pâncreas. Estudos imunoistoquímicos mostraram marcação em ductos coletores renais e porção ascendente espessa da alça de Henle, em epitélios ductais biliar e pancreático e, no período embrionário, em broto ureteral, ductos biliar e pancreático e glândula salivar. Análises por imunofluorescência e microscopia imunoeletrônica sugerem que poliductina se localize em cílio apical primário, membrana apical e citoplasma de células do ducto coletor. Nossos resultados indicam que PKHD1 codifica isoformas de membrana e solúveis
Title in English
Analysis of the expression pattern of the PKHD1 gene product, mutated in autossomal recessive polycystic kidney disease
Keywords in English
BLOTTING WESTERN/methods
IMMUNOHISTOCHEMISTRY/methods
KIDNEY TUBULES COLLECTING/pathology
KIDNEY TUBULES COLLECTING/physiopathology
MEMBRANE PROTEINS/analysis
MICROSCOPY FLUORESCENCE/methods
MICROSCOPY IMMUNOELECTRON/methods
POLYCYSTIC KIDNEY AUTOSOMAL RECESSIVE/etiology
POLYCYSTIC KIDNEY AUTOSOMAL RECESSIVE/genetics
POLYCYSTIC KIDNEY AUTOSOMAL RECESSIVE/physiopathology
PROTEIN ISOFORMS/analysis
Abstract in English
PKHD1, the gene mutated in autosomal recessive polycystic kidney disease, presents a complex splicing pattern, associated with multiple alternative transcripts. In this work we have studied the expression profile of its product, polyductin. Western blot analysis revealed putative membrane products of > 440 kDa and 230 kDa, and of ~140 kDa in soluble fractions in kidney, liver and pancreas. Immunohistochemistry studies showed staining in renal collecting duct and thick ascending limb of Henle, in biliary and pancreatic ductal epithelia and, in the embryonic period, in ureteric bud, biliary and pancreatic ducts and salivary gland. Immunofluorescence and immunoelectron microscopy studies suggest that polyductin localizes to primary apical cilium, apical membrane and cytoplasm of collecting duct cells. Our data indicate that PKHD1 codifies membrane and soluble isoforms
 
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Publishing Date
2005-10-05
 
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