bromochloroacetic-acid and nephrin

Several podocyte-related markers are organized to express in glomerular differentiation. However, whether expression of them is virtually synchronized and a reliable indicator of the state of differentiation is unknown. The present study investigated, by immunohistochemistry, the divergent expression of several podocyte markers in the improperly differentiated glomeruloid bodies from four cases of Wilms tumors. The glomeruloid bodies were classified into immature (IGB) or mature forms (MGB) based on morphology and epithelial features. Podocytes in IGB expressed WT1, synaptopodin, podocalyxin, and nephrin, and their expression was stronger in MGB. In contrast, Pax2 was strong in IGB and diminished in MGB. p27 was first expressed in MGB. The expression pattern in each molecule mimics normal glomerulogenesis. Podocytes in MGB showed persistent expression of bcl-2 and cytokeratin with synaptopodin, podocalyxin, and nephrin by serial section, a finding unusual for normal glomerulogenesis. Moreover, parietal cells in MGB also occasionally expressed these podocyte markers. The ultrastructure revealed that podocytes in MGB showed tight junctions without foot process formations, which indicated incomplete differentiation. These results suggest that a set of podocyte differentiation markers are occasionally diversely expressed, and raise the possibility that expression of these markers is insufficient to determine the state of terminal differentiation in podocytes.

Topics: Biomarkers, Tumor; Female; Humans; Infant; Keratins; Kidney Glomerulus; Kidney Neoplasms; Male; Membrane Proteins; Microfilament Proteins; Muscle Proteins; Proteins; Proto-Oncogene Proteins c-bcl-2; Sialoglycoproteins; Tight Junctions; Wilms Tumor; WT1 Proteins

The NPHS1 gene product, nephrin, is a crucial component of the glomerular filtration barrier preventing proteinuria and previously assumed to be kidney-specific. The aim of this study was to describe the expression of nephrin mRNA and protein in human pancreas as well as identify the nephrin-expressing cell types.. RNA dot blot, reverse transcriptase-polymerase chain reaction, sequencing, immunoblotting and dual immunofluorescence were used for the characterisation of nephrin in the pancreas.. Except for the kidney, the pancreas was found to be the only tissue expressing nephrin as screened with a human tissue RNA dot blot. The expression was verified with reverse transcriptase-polymerase chain reaction and by sequencing nephrin from a human pancreatic complementary DNA library. Nephrin antibody in immunoblot detected a 165,000 M(r) protein in the pancreas. Dual immunofluorescence showed that nephrin was specifically localised in the beta cells of the islets of Langerhans. There was no overlap with glucagon, somatostatin, or the ductal cell marker cytokeratin 19.. These data show that nephrin is a novel molecule of pancreatic beta cells.

Topics: Cadaver; Fluorescein-5-isothiocyanate; Fluorescent Antibody Technique; Fluorescent Dyes; Gene Expression; Glucagon; Humans; Immunoblotting; Islets of Langerhans; Keratins; Kidney; Kidney Cortex; Membrane Proteins; Molecular Weight; Pancreas; Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Somatostatin