Project description:Neural activity-dependent translation is essential for synaptic plasticity and diverse brain functions. Translation involves not only canonical main open reading frames (mORFs) but also upstream ORFs (uORFs), which may regulate mORF expression. However, due to technical limitations, systematic investigation of activity-dependent uORFs and mORFs in brain tissues remains challenging. Here, we developed a ribosome tagging and purification strategy that bypasses the prolonged turnover of ribosomal proteins, enabling ribosome profiling with one-hour temporal resolution after neural stimulation. Applying this strategy to mouse hippocampal slices undergoing long-term potentiation, we identified hundreds of activity-induced mORFs and uORFs, a subset of which served as robust markers for activity status. Notably, ~22% of the upregulated uORFs overlapped with those induced by the integrated stress response, suggesting potential crosstalk between these signaling pathways. This study provides a useful technique and resources for deciphering molecular mechanisms underlying activity and translation-dependent brain functions in health and disease.
Project description:Silkworms show a reproductive behavior induced by sex pheromone. To elucidate the neral mechanism of sex pheromone induced sexual behavior in the silkworm, we attempted to use the neural activity-induced gene as a neural activity marker. Since no neural activity-induced gene was identified in the silkworm, we conducted screening of neural activity-induced gene using the male silkworm brain. By the screening, we identified Bhr38 as a novel neural activity-induced gene, and succeded to comprehensively map the active neruons in the silkworm brain in response to the sex pheromone exposure. Further, we found that Dhr38, the Drosophila homologue of Bhr38, also expressed in a neural activity dependent manner. These results strongly suggest that Hr38 is a highly conserved neural activity-induced gene. The male silkworms were exposed to the female odor for 30 min (group P). Non-treated male silkworms were used as the control group group C. Ten brains were collected for each sample and stored at -80°C until use. Total RNA was isolated by the TRIzol reagent and subjected to microarray experiments using the custam made (8x16k) Oligo Microarray (Agilent Technologies, Inc.).
Project description:Silkworms show a reproductive behavior induced by sex pheromone. To elucidate the neral mechanism of sex pheromone induced sexual behavior in the silkworm, we attempted to use the neural activity-induced gene as a neural activity marker. Since no neural activity-induced gene was identified in the silkworm, we conducted screening of neural activity-induced gene using the male silkworm brain. By the screening, we identified Bhr38 as a novel neural activity-induced gene, and succeded to comprehensively map the active neruons in the silkworm brain in response to the sex pheromone exposure. Further, we found that Dhr38, the Drosophila homologue of Bhr38, also expressed in a neural activity dependent manner. These results strongly suggest that Hr38 is a highly conserved neural activity-induced gene.
Project description:Neuronal activity-dependent gene expression plays important roles in neural plasticity. We use electroconvulsive stimulation (ECS) as an in vivo model for neuronal activation to identify genes that are regulated by neuronal activity. Dentate gyri (DG) were microdissected 4 hours after sham or ECS treatment for gene expression profiling. 4 total samples were analysed (2 for each condition). Averaged expression values between sham and ECS samples were pair-wise compared.
Project description:Neuronal activity-dependent gene expression plays important roles in neural plasticity. We use electroconvulsive stimulation (ECS) as an in vivo model for neuronal activation to identify genes that are regulated by neuronal activity. Dentate gyri (DG) were microdissected 4 hours after sham or ECS treatment for gene expression profiling.
Project description:Human induced pluripotent stem cells (hiPSCs) have emerged as a promising in vitro model system for studying neurodevelopment. However, current models remain limited in their ability to incorporate tunable biochemical and biomechanical signaling cues imparted by the neural extracellular matrix (ECM). The native brain ECM is viscoelastic and stress-relaxing, exhibiting a time-dependent response to an applied force. To recapitulate the remodelability of the neural ECM, we developed a family of protein-engineered hydrogels crosslinked with either static or dynamic covalent bonds that exhibit tunable stress relaxation rates. hiPSC-derived neural progenitor cells (NPCs) encapsulated within these gels underwent relaxation rate-dependent maturation. Specifically, NPCs within hydrogels with faster stress relaxation rates extended longer, more complex neuritic projections, exhibited decreased metabolic activity, and expressed higher levels of genes associated with neural maturation. By inhibiting actin polymerization, we observed decreased neuritic projections and a concomitant decrease in the expression of neural maturation genes. Taken together, these results suggest that microenvironmental viscoelasticity is sufficient to bias human NPC maturation.