Single molecule fluorescence in situ hybridisation for quantitating post-transcriptional regulation in Drosophila brains.

ABSTRACT: RNA in situ hybridization is a powerful method to investigate post-transcriptional regulation, but analysis of intracellular mRNA distributions in thick, complex tissues like the brain poses significant challenges. Here, we describe the application of single-molecule fluorescent in situ hybridization (smFISH) to quantitate primary nascent transcription and post-transcriptional regulation in whole-mount Drosophila larval and adult brains. Combining immunofluorescence and smFISH probes for different regions of a single gene, i.e., exons, 3'UTR, and introns, we show examples of a gene that is regulated post-transcriptionally and one that is regulated at the level of transcription. Our simple and rapid protocol can be used to co-visualise a variety of different transcripts and proteins in neur

SUBMITTER: Yang L 

PROVIDER: S-EPMC5595163 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature