Profiling local translatomes and RNA binding proteins of somatosensory neurons reveals specializations of individual axons [TRAP-seq]

Individual neurons have one or more axons that often extend long distances and traverse multiple microenvironments, yet it is not known how the composition of individual axons is established or locally modulated to enable neuronal plasticity. Here, we use spatial translatomics to identify local axonal translatomes in anatomically and functionally specialized neurons in the dorsal root ganglia (DRG). DRG neurons extend long central and peripheral axons in separate directions and through distinct microenvironments to enable somatosensation. Using Translating Ribosome Affinity Purification and RNA sequencing, we generated a comprehensive resource of mRNAs preferentially translated within each axon. Locally translated proteins include pain receptors, ion channels, and translational machinery, which establish distinct electrophysiologic properties and regenerative capacities for each axon. Furthermore, we identify RNA-binding proteins associated with sorting and transporting functionally related mRNAs. These findings provide resources for addressing how axonal translation shapes the spatial organization of neurons and enables subcellular neuroplasticity. Overall design: Translating Ribosome Affinity Purification (TRAP) paired with RNA-seq profiling of central axons, peripheral axons, and cell somas of Nav1.8-positive dorsal root ganglia (DRG) sensory neurons. Central axons were collected from lumbar 1-6 dorsal roots and spinal cord, peripheral axons were collected from sciatic nerves, and cell somas were collected from lumbar 1-6 dorsal root ganglia.