Spatially Precise In Situ Transcriptomics in NPF neurons
2The Alexander Kofkin Faculty of Engineering, Bar Ilan, Israel
3The Multidisciplinary Brain Research Center, Bar Ilan, Israel
NPF neurons is a subset of neurons in Drosophila brain that express neuropeptide F, they share similarities with the vertebrate neuropeptide Y. These neurons promote behaviors that are required for survival such as, feeding and mating. Previous work in this filed are based on all brain RNA extraction or bulk sequencing. These methods are limited from spatial resolution and ability to focus on a single cell. The ability to identify the spatial location of transcripts whether in the soma, dendrites, synapses, or axons is crucial for the understanding of molecular processes that leads to different behaviors, such as motivation, memory, learning and more.
In this project we used in situ expansion sequencing approach that combined genomics and microscopy, enabling the identification of individual RNA molecules with nanometer resolution in intact tissues. We succeed to identify spatial location of selected transcripts of NPF neurons. This is possible due to expansion of the brain and Illumina sequencing. We wish to expand the identification to 100 genes and examine the impact of different behaviors on the localization of the genes in NPF. Using this approach, we hope to be able to classify molecular changes as a function of social experience of the fruit fly.
This groundbreaking project examines for the first time the spatial location of genes expressed in neurons at high nanometer resolution. Special emphasis will be placed on transcripts located in synapses and dendrites, as well as differences in the expression pattern of the population of NPF neurons.