Project description:To investigate if Fibcd1 mediates CSPG signalling to neurons, we established a primary mouse hippocampal neuron culture plated +/- a coating of CSPGs. After determining that Fibcd1 KO neurons are partially resistant to CSPG signals, we sought to identify the transcriptional finger print of CSPG-Fibcd1 signalling by nulk RNA-seq.
Project description:The experiment was performed to identify autophagy targets in wildtype and autophagy-deficient primary neurons. Therefore, cortico-hippocampal neurons were isolated from Atg5flox:CAG-CreTmx mice and treated with Tamoxifen (Tmx) to induce the knockout (KO) or EtOH for wildtype (WT) in-vitro. WT and KO Neurons were harvested at day in-vitro (DIV) 15-16, and global proteome analysis was measured by LC-MS/MS.
Project description:The experiment was performed to identify autophagy targets in wildtype and autophagy-deficient primary neurons. Therefore, cortico-hippocampal neurons were isolated from Atg16L1flox:CAG-CreTmx mice and treated with Tamoxifen (Tmx) to induce the knockout (KO) or EtOH for wildtype (WT) in-vitro. WT and KO Neurons were harvested at day in-vitro (DIV) 15-16, and global proteome analysis was measured by LC-MS/MS.
Project description:We conducted RNA-Seq (using Direct Ligation of Adapters to first strand cDNA) in neurons from E16.5 mouse embryonic cortices from WT and SMCX KO mice and harvested after 10 days in vitro culture. We sequenced RNA samples after 10 days in vitro cultures in biological and technical duplicates. 4 RNA samples from WT and SMCX KO neurons. We also sequenced RNA samples from same neurons after stimulation with KCl for 60 mins. So a total of 8 RNA-Seq samples.
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:Despite their widespread use as a research model, a comprehensive, quantitative proteomic profile of the cultured hippocampal neurons has remained unexplored. Here, we provide the first global proteomic characterization of primary murine hippocampal neurons cultured for 14 days under near-physiological glucose conditions (2.5 mM). Using label-free mass spectrometry and the Total Protein Approach (TPA), we quantified over 5,500 proteins, revealing a metabolic signature consistent with oxidative phosphorylation and a neuron-specific proteome largely devoid of astrocytic markers. Unexpectedly, glial fibrillary acidic protein (GFAP) was highly abundant in pure neuronal cultures, and its knockdown impaired neuronal viability and morphology. These findings suggest a compensatory role for GFAP in neurons cultured without astrocytic support. Additionally, our data highlight significant underexpression of key synaptic proteins compared to hippocampal tissue, underscoring the limitations of neuron-only culture systems. This study establishes a new molecular reference for hippocampal neurons and offers insights into neuronal adaptation in simplified environments.
Project description:The experiment was performed to identify autophagy targets in wildtype and autophagy-deficient forebrain excitatory neurons. Therefore, neurons were isolated from the cortex, hippocampus and striatum of 2-3 weeks old Atg5flox/flox:CamKIIα-Cretg/wt:tdTomato+ (KO) and Atg5wt/wt:CamKIIα-Cretg/wt:tdTomato+ (WT) mice. Neurons in suspension were FACS sorted and excitatory forebrain neurons expressing tdTomato were forwarded to global proteome analysis assessed by LC-MS/MS.
Project description:The experiment was performed to identify autophagy targets in wildtype and autophagy-deficient forebrain inhibitory neurons. Therefore, neurons were isolated from the cortex, hippocampus and striatum of 2-3 weeks old Atg5flox/flox:Slc32a1-Cretg/wt:tdTomato+ (KO) and Atg5wt/wt:Slc332a1-Cretg/wt:tdTomato+ (WT) mice. Neurons in suspension were FACS sorted and inhibitory forebrain neurons expressing tdTomato were forwarded to global proteome analysis assessed by LC-MS/MS.
Project description:Poly(A) RNA profiling upon Gld2 knockdown in cultured hippocampal neurons Neurons transduced with scrambled and Gld2 knowdown shRNA