Detection of multiple vascular endothelial growth factor splice isoforms in single glomerular podocytes

摘要: Detection of multiple vascular endothelial growth factor splice isoforms in single glomerular podocytes. Glomerular podocytes are major determinants filtration permselectivity the glomerulus. Although molecular mechanisms determining characteristics unit incompletely understood, (VEGF) has been implicated. To analyze this process situ, we established a method that allows exploration vivo mRNA expression using single-cell reverse transcriptase-polymerase chain reaction (RT-PCR). Microdissected mouse glomeruli were held patch-clamp apparatus, and harvested by aspiration. After lysis, cells transcribed, PCR was performed (45 cycles). The podocyte nature material confirmed detection podocyte-specific (glomerular epithelial protein 1 Wilms' tumor 1). Using specific oligonucleotide primers, VEGF detected obtained from renal cortex, microdissected glomeruli, cultured murine podocytes, situ. All examined expressed three (121, 165, 189). These differ their capacity for binding to extracellular matrix could have different potencies regulating permeability. Our approach should allow semiquantitative, isoform-specific evaluation during nephrogenesis disease.