Transcription profiling of mouse FACS-purified cortical projection neurons
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ABSTRACT: This SuperSeries is composed of the following subset Series:; GSE2039: FACS purified cortical projection neurons; GSE17783: Analysis of gene expression in FACS-purified cortical projection neurons using Affymetrix 430 2.0 microarrays Experiment Overall Design: Refer to individual Series
Project description:3 subtypes of cortical projection neurons were purified by fluorescence-activated cell sorting (FACS) at 4 different stages of development from mouse cortex. A detailed description of the data set is described in Arlotta, P et al (2005) and Molyneaux, BJ et al (2009). The hybridization cocktails used here were originally applied to the Affymetrix mouse 430A arrays and submitted as GEO accession number GSE2039. The same hybridization cocktails were then applied to the Affymetrix mouse 430 2.0 arrays, and those data are contained in this series. Experiment Overall Design: Three subtypes of cortical neurons were purified by FACS at multiple stages of mouse brain development. The neuron subtypes are: corticospinal motor neurons (CSMN), callosal projection neurons (CPN), and corticotectal projection neurons (CTPN). The stages of development included embryonic day 18 (E18), postnatal day 3 (P3), postnatal day 6 (P6), and postnatal day 14 (P14). CSMN and CPN were analyzed at all four stages, while CTPN were only analyzed at P14. The replicates included in the data set are all true biological replicates with independent sample collection for each.
Project description:3 subtypes of cortical projection neurons were purified by fluorescence-activated cell sorting at 4 different stages of development from mouse cortex. A detailed description of the data set is described in Arlotta, P et al (2005)
Project description:Molecular mechanisms controlling specification and differentiation of distinct neuron subtypes in the cerebral cortex are not well understood. Corticothalamic projection neurons (CThPN) are a diverse set of neurons, critical for function of the neocortex, but little is known about the molecular mechansims controlling their development. We used microarrays to detail CThPN gene expression at 3 developmental time points. We then compared CThPN gene expression, with the gene expression of other neuron subtypes in the cerebral cortex at the same stages (previously described in Arlotta et a., 2005) CThPN were retrogradely label with fluorescent microespheres injected into the thalamus, and were FACS purified at 3 developmental stages for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Amyotrophic Lateral Sclerosis is clinically defined as the combined degeneration of the corticospinal and corticobulbar neurons (CSN) along with the bulbar and spinal motor neurons (SMN). While a growing body of evidence points to the motor cortex, where CSN are located, as the potential initiation site of ALS, little is known about how CSN degenerate. To gain insights into the molecular mechanisms behind CSN selective degeneration, we first developed an approach to purify this neuronal population from the cerebral cortex of adult wild-type and Sod1G86R mice, combining retrograde labelling and Fluorescence Activated Cell Sorting. In parallel, a second population of cortical neurons, the callosal projection neurons (CPN) located in the layers II/III of the cerebral cortex were also purified. CPN and CSN are both cortical excitatory projection neurons but as opposed to CSN, CPN do not degenerate in Sod1G86R mice, and served as control population. CSN and CPN were purified from same animals, at two presymptomatic ages (3 and 60 days) and two symptomatic ages, 90 and 105 days. A total of 57 samples were further processed and analysed (CSN: 30d: n=4 WT and 3 Sod1G86R; 60d: n=4 WT and 4 Sod1G86R; 90d: n=4 WT and 4 Sod1G86R; 105d: n=4 WT and 2 Sod1G86R; CPN: 30d: n=4 WT and 3 Sod1G86R; 60d: n=2 WT and 3 Sod1G86R; 90d: n=4 WT and 4 Sod1G86R; 105d: n=4 WT and 4 Sod1G86R).
Project description:3 subtypes of cortical projection neurons were purified by fluorescence-activated cell sorting at 4 different stages of development from mouse cortex. A detailed description of the data set is described in Arlotta, P et al (2005). Keywords = corticospinal motor neuron callosal corticotectal cortex development FACS
Project description:Corticospinal motor neurons (CSMN) are one specialized class of cortical excitatory neurons, which connect layer Vb of the cortex to the spinal cord. a master transcription factor –Forebrain expressed zinc finger 2 (Fezf2) – has been identified that is necessary for the fate specification of CSMN. Fezf2 alone can cell-autonomously instruct the acquisition of CSMN-specific features when expressed in diverse, permissive cellular contexts, in vivo. In order to understand the molecular logic underlying the acquisition of CSMN traits upon Fezf2 expression, we compared the in vivo gene expression of FACS-purified cortical progenitors that ectopically expressed Fezf2 to control progenitors. We used in utero electroporation to deliver Fezf2GFP or CtrlGFP expression vectors to neocortical progenitors at E14.5, when they primarily generate CPN of the upper layers. Overexpression of Fezf2 in these progenitors is sufficient to instruct a fate-switch resulting in the generation of CSMN and other subtypes of corticofugal projection neurons. Fezf2GFP- and CtrlGFP -electroporated progenitors were FACS-purified at 24 and 48 hours after surgery and acutely profiled by microarrays.
Project description:Single-cell multiomic techniques have sparked immense interest in developing a comprehensive multi-modal map of diverse neuronal cell types and their brain-wide projections. However, investigating the complex wiring diagram, spatial organization, transcriptional, and epigenetic landscapes of brain-wide projection neurons is hampered by the lack of efficient and easily adoptable tools. Here we introduce Projection-TAGs, a retrograde AAV platform that allows multiplex tagging of projection neurons using RNA barcodes. By using Projection-TAGs, we performed multiplex projection tracing of the mouse cortex and high-throughput single-cell profiling of the transcriptional and epigenetic landscapes of the cortical projection neurons. Projection-TAGs can be leveraged to obtain a snapshot of activity-dependent recruitment of distinct projection neurons and their molecular features in the context of a specific stimulus. Given its flexibility, usability, and compatibility, we envision that Projection-TAGs can be readily applied to build a comprehensive multi-modal map of brain neuronal cell types and their projections.
2025-04-01 | GSE277718 | GEO
Project description:Gene expression in FACS-purified cortical projection neurons
Project description:Cortical projection neurons comprise classes of neurons connecting the cerebral cortex to distant targets in the nervous system. Each projection neuron class acquires a distinct molecular identity and develops characteristic patterns of axonal projection and dendritic arborization that determine their inputs, outputs, and distinct functions. How different aspects of neuron identity, including axonal and dendritic morphology, are coordinated appropriately in each specific neuron type is not known. A network of transcription factors, including the selector gene Fezf2, is central to specifying cortical projection neuron fates. However, regulation down-stream of these fate-determinant transcription factors to control different aspects of neuron identity is not understood, particularly as it relates to the development of distinct dendritic arbors that determine the inputs to each projection neuron class. Here we show that the miR-193b~365 microRNA cluster downstream of Fezf2 cooperatively represses the signaling molecule Mapk8 to regulate dendritic development in a neuron subtype-specific manner.