Cell Biological Mechanisms of Activity-Dependent Synapse to Nucleus Translocation of CRTC1 in Neurons

Cell Biological Mechanisms of Activity-Dependent Synapse to Nucleus Translocation of CRTC1 in Neurons

Blog Article

Previous studies have revealed a critical role LEDs for CREB-regulated transcriptional coactivator (CRTC1) in regulating neuronal gene expression during learning and memory.CRTC1 localizes to synapses but undergoes activity-dependent nuclear translocation to regulate the transcription of CREB target genes.Here we investigate the long-distance retrograde transport of CRTC1 in hippocampal neurons.

We show that local elevations in calcium, triggered by activation of synaptic glutamate receptors and L-type voltage-gated calcium channels, initiate active, dynein-mediated retrograde transport of CRTC1 along microtubules.We identify a nuclear localization signal within CRTC1, and characterize three conserved serine residues whose dephosphorylation is required for nuclear import.Domain analysis reveals that the amino-terminal third of CRTC1 contains all of the Goldenrod signals required for regulated nucleocytoplasmic trafficking.

We fuse this region to Dendra2 to generate a reporter construct and perform live-cell imaging coupled with local uncaging of glutamate and photoconversion to characterize the dynamics of stimulus-induced retrograde transport and nuclear accumulation.