Detection of Tissue Kallikrein and Kinin B1 and B2 Receptor MRNAs in Human Brain by in Situ RT-PCR

Tissue kallikrein (TK) and kinin receptors have been immuno-localized in various areas of the human nervous system, suggesting that the kallikrein-kinin system (KKS) may be functionally active in the brain. The aim of this study was to determine the cellular expression of TK and kinin B1 and B2 receptor mRNAs in specific regions of the human brain by in situ reverse transcriptase polymerase chain reaction. Autopsy samples of the brain, spinal cord, kidney and salivary gland were embedded in paraffin. Sections (5 microm), adhered onto silane coated glass slides, were treated with Proteinase K and DNase, followed by reverse transcription polymerase chain reaction with specific KKS primers and digoxigenin-dUTP. Detection of the digoxigenin-label demonstrated localization of TK, B1 and B2 mRNAs in the cytoplasm of some neuronal cell bodies in the hypothalamus, thalamus, frontal cortex and spinal cord. TK mRNA was also observed in the ependymal cells lining the cerebral ventricles and epithelial cells of the choroid plexus. In the choroid plexus, only B1 gene expression was observed in some choroidal epithelial cells while no B2 labeling was detected. The identification of mRNAs to TK, B1 and B2 kinin receptors in human nervous tissue supports previous evidence for the presence of the KKS in the brain and confirms localized protein synthesis.