Project description:To identify cAMP-dependent lncRNAs, we treated DIV16 primary hippocampal neurons with 50uM Forskolin or equal volume DMSO for 30 minutes prior to RNA isolation using Trizol and Total RNA sequencing. We submitted samples to Scripps Florida Genomics Core for library preparation and sequencing.
Project description:To identify cAMP-dependent and mGluR1/5-dependent RNAs, we treated DIV16 primary hippocampal neurons with 50uM Forskolin, 30uM DHPG, or equal volume DMSO for 30 minutes prior to RNA isolation using Trizol and Total/small RNA sequencing. We submitted samples to Scripps Florida Genomics Core for library preparation and sequencing.
Project description:Analysis of T3 and CORT dependent gene expression tested in the mouse hippocampal neuronal cell line HT-22. The experiment was done to identify genes which are differentially regulated by the two hormones as well as to identify genes which are synergistically regulated by combined T3 + CORT treatment. Results provide an exploratory dataset for studying T3 and CORT regulation in mouse hippocampal neurons.
Project description:We induced over-expression and under-expression of Camk2b in cultured rat hippocampal neurons through transfection with lentivirus plasmids. Then isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomics followed by bioinformatics analyses were carried out to explore the impacts of Camk2b dysexpression on the proteome of the neurons.
Project description:To assess neuronal expression divergence between mice and rats, we used the Affymetrix array platform to assay the transcriptomes of micro-dissected individual soma and pool of dendrites of hippocampal neurons in dispersed primary cell cultures from rat and mouse. Using microdissected soma and dendrites from primary cultures of hippocampal neurons of two mouse strains (C57BL/6 and Balb/c) and one rat strain (Sprague-Dawley), we investigate via microarrays, subcellular localization of mRNAs in neurons
Project description:To investigate the possible roles on mitotic/microtubule-related genes during hippocampal neuron differentiation, we dissected E18.5 mouse embryonic hippocampi, cultured neurons in vitro and collected total RNA samples from different timepoints of differentiation.
Project description:Ongoing neuronal activity during development and plasticity acts to refine synaptic connections and contributes to the induction of plasticity and ultimately long term memory storage. Activity-dependent post-transcriptional control of mRNAs occurs through transport to axonal and dendritic compartments, local translation, and mRNA stability. We have identified a mechanism that contributes to activity-dependent regulation of mRNA stability during synaptic plasticity. In this study we demonstrate rapid, post-transtriptional control over process-enriched mRNAs by neuronal activity. Systematic analysis of the 3'-UTRs of destablized transcripts, identifies enrichment in sequence motifs corresponding to miRNA binding sites. The miRNAs that were identified, miR-326-3p/miR-330-5p, miR-485-5p, miR-666-3p, and miR-761 are predicted to regulate networks of genes important in plasticity and development. We find that these miRNAs are developmentally regulated in the hippocampus, many increasing by postnatal day 14. We further show that miR-485-5p controls NGF-induced neurite outgrowth in PC12 cells, tau expression, and axonal development in hippocampal neurons. miRNAs can function at the synapse to rapidly control and affect short- and long-term changes at the synapse. These processes likely occur during refinement of synaptic connections and contribute to the induction of plasticity and learning and memory. 12 hippocampal cell culture samples analysed, 3 coverslips pooled per sample. Treatments are as follows: Block: an inhibitor cocktail containing 50 µM D-APV, 40 µM CNQX, and 100 nM TTX for 3 hrs Activity: 50 µM bicuculline (BiC)/500 µM 4-Aminopyridine ActD: 25 µM actinomycin D
Project description:Gene expression profiles of Ngn3-overexpressing cultured hippocampal neurons was compared to the profile of the corresponding control populations. (neurons expressing GFP). Neurogenin3, a proneural transcription factor controlled by Notch receptor, is involved in hippocampal neuron differentiation and synapses, but little is known about the molecular bases of Ngn3 activity in neurons. Microarray analysis indicated that overexpression of Ngn3 upregulated a number of genes related with cytoskeleton dynamics. One of then was Fmn1 whose protein is associated with actin and microtubule cytoskeleton. Overexpression of the isoform Fmn1-Ib in cultured hippocampal neurons induced an increase in the number of primary dendrites and in the number of glutamatergic synaptic inputs without affecting GABAergic synapses resulting in a modification in the balance between excitation and inhibition. The same changes were provoked by overexpression of Ngn3. In addition downregulation of Fmn1 by the use of Fmn1-siRNAs impaired such morphological and synaptic changes induced by Ngn3 overexpression in neurons. These results reveal a previously unknown involvement of Formin1 in dendritic and synaptic plasticity as a key protein in the Nng3 signaling pathway that contributes to understanding of molecular mechanisms of the neuronal differentiation. Cultured hippocampal neurons were transduced using Sindbis virus bearing myc-tagged Ngn3 or GFP as control. Cells were lysed and total RNA was extracted.Gene expression profiles were obtained for each sample and compared
Project description:Poly(A) RNA profiling upon Gld2 knockdown in cultured hippocampal neurons Neurons transduced with scrambled and Gld2 knowdown shRNA
Project description:To assess neuronal expression divergence between mice and rats, we used the Affymetrix array platform to assay the transcriptomes of micro-dissected individual soma and pool of dendrites of hippocampal neurons in dispersed primary cell cultures from rat and mouse. Using microdissected soma and dendrites from primary cultures of hippocampal neurons of two mouse strains (C57BL/6 and Balb/c) and one rat strain (Sprague-Dawley), we investigate via microarrays, subcellular localization of mRNAs in neurons