[2] In situ hybridization techniques to map processing enzymes

摘要: Publisher Summary This chapter discusses the in situ hybridization techniques to map processing enzymes. In have been consistently improved over last decade, enabling one study expression of low-abundance messages at cellular level, such as RNA transcripts encoding With assistance polymerase chain reaction technology and molecular biological procedures, cRNA probes labeled with radioisotopes proved be most sensitive tools, reaching a detection limit few molecules per cell, compromised only by background labeling. Applying this mapping studies enzymes, determination region-specific enzyme sets has given new insights into tissue-specific differences posttranslational elucidated role some these enzymes for specific precursors. Research community is able identify prohormone convertases associated predominantly secretory pathway secretion neurons endocrine cells, ubiquitously expressed. An important lesson learned from that not presence or absence convertase but relative ratios crucial tissue- cell-specific processing. Using nonradioactive it shown can now carried out high anatomical resolution single-cell level.