Click-iT method can be used to trace the neurons where the newly synthesized RNA transcripts occur. Our experiments performed with the CNS of terrestrial mollusk Helix demonstrate that 5-ethynyluridine (EU) is selectively incorporated in RNA but not in DNA. The time of EU accumulation necessary for its detection was about several hours. EU was injected into the body cavity of adult mollusks, and was detectable in neurons for several days. In juveniles, EU was introduced via bathing of snails in the EU-containing saline, and was reliably detected within time period of several weeks. Our data suggest that short-living forms of RNA cannot be detected by Click-iT method, while the long-living forms of RNA can be spatially detected in individual neurons.
Keywords: 5-ethynyluridine; Click-iT reaction; RNA; mollusks; neurons.