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CUT&RUN of Sox2 binding in the mouse P0 visual thalamus (dLGN)

ABSTRACT: CUT&RUN of Sox2 binding in the mouse P0 visual thalamus (dLGN)

PROVIDER: PRJEB62553 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR11475105_1.fastq.gz	Fastqsanger.gz
	ERR11475105_2.fastq.gz	Fastqsanger.gz
	ERR11475106_1.fastq.gz	Fastqsanger.gz
	ERR11475106_2.fastq.gz	Fastqsanger.gz
	ERR11475107_1.fastq.gz	Fastqsanger.gz

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Effect of thalamic deletion of mouse Sox2, or Nr2f1, on gene expression in the visual thalamus (dorsolateral geniculate nucleus, dLGN)

Project description:Effect of thalamic deletion of mouse Sox2, or Nr2f1, on gene expression in the visual thalamus (dorsolateral geniculate nucleus, dLGN), as described in "SOX2 and NR2F1 coordinate the gene expression program of the early postnatal visual thalamus" (bioRxiv doi number to be updated) To address the effect of Sox2, or Nr2f1, Cre-mediated thalamic deletion in mouse, on gene expression in the visual thalamus (dLGN), we dissected the dLGN from mutant (Sox2, or Nr2f1) and control mice at postnatal day 0 (P0), for both Sox2 and Nr2f1 mutants. We then performed gene expression analysis by RNAseq.

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CUT&RUN of Sox2 binding in the mouse P0 visual thalamus (dLGN)

Project description:CUT&RUN was performed for Sox2 on ex-vivo dissected visual thalamic nuclei from P0 mice, revealing context specific activity of Sox2 binding in differentiated neurons.

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Effect of thalamic deletion of mouse Sox2, or Nr2f1, on gene expression in the visual thalamus (dorsolateral geniculate nucleus, dLGN)

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Project description:Analysis of thalamus and hypothalamus under conditions of visual deprivation by dark-rearing (DR). Animals subjected to DR from birth till postnatal day (P) 14. Results provide insight into the role of visual inputs in the regulation of gene expression in thalamus and hypothalamus during development. RNA sample was taken from thalamus and hypothalamus of 14-day-old mouse reared in standard or dark condition. Comparisons among groups were made by one-color method with normalized data from Cy3 channels for data analysis.

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Single-cell and single-nucleus RNA-seq uncovers shared and distinct axes of variation in dorsal LGN neurons in mice, non-human primates, and humans

Project description:Abundant evidence supports the presence of at least three distinct types of thalamocortical (TC) neurons in the primate dorsal lateral geniculate nucleus (dLGN) of the thalamus, the brain region that conveys visual information from the retina to the primary visual cortex (V1). Different types of TC neurons in mice, humans, and macaques have distinct morphologies, distinct connectivity patterns, and convey different aspects of visual information to the cortex. To investigate the molecular underpinnings of these cell types, and how these relate to differences in dLGN between human, macaque, and mice, we profiled gene expression in single nuclei and cells using RNA-sequencing. These efforts identified four distinct types of TC neurons in the primate dLGN: magnocellular (M) neurons, parvocellular (P) neurons, and two types of koniocellular (K) neurons. Despite extensively documented morphological and physiological differences between M and P neurons, we identified few genes with significant differential expression between transcriptomic cell types corresponding to these two neuronal populations. Likewise, the dominant feature of TC neurons of the adult mouse dLGN is high transcriptomic similarity, with an axis of heterogeneity that aligns with core vs. shell portions of mouse dLGN. Together, these data show that transcriptomic differences between principal cell types in the mature mammalian dLGN are subtle relative to the observed differences in morphology and cortical projection targets. Finally, alignment of transcriptome profiles across species highlights expanded diversity of GABAergic neurons in primate versus mouse dLGN and homologous types of TC neurons in primates that are distinct from TC neurons in mouse.

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Project description:Nuclei of the mammalian thalamus are aggregations of neurons with unique architectures and input-output connections, yet the molecular determinants of their organizational specificity remain unknown. By comparing expression profiles of thalamus and cerebral cortex in adult rhesus monkeys we identified transcripts that are unique to dorsal thalamus or to individual nuclei within it. Real-time quantitative polymerase chain reaction and in situ hybridization analyses confirmed the findings. Expression profiling of individual nuclei microdissected from the dorsal thalamus revealed additional subsets of nucleus-specific genes. Functional annotation using Gene Ontology (GO) vocabulary and Ingenuity Pathway analysis revealed over-representation of GO categories related to development, morphogenesis, cell-cell interactions, and extracellular matrix within the thalamus- and nucleus-specific genes-many involved in the Wnt signaling pathway. Examples included the transcription factor TCF7L2, localized exclusively to excitatory neurons, a calmodulin-binding protein PCP4, the bone extracellular matrix molecules SPP1 and SPARC, and other genes involved in axon outgrowth and cell matrix interactions. Other nucleus-specific genes such as CBLN1 are involved in synaptogenesis. The genes identified likely underlie nuclear specification, cell phenotype and connectivity during development and their maintenance in the adult thalamus. Experiment Overall Design: To determine the molecular underpinnings of nuclear specificity in the dorsal thalamus we isolated micro-punches of tissue from nucleus-specific regions and processed them for microarray analysis. Replicate samples from 5 separate dorsal thalamic nuclei were processed and compared to identify genes unique to each region. Affymetrix U133A Gene Chips were used. All of the samples were isolated from untreated adult monkey brain.

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Guidance landscapes unveiled by quantitative proteomics to control reinnervation in adult visual system

Project description:Long-distance regeneration of the central nervous system (CNS) has been achieved from the eye to the brain through activation of neuronal molecular pathways or pharmacological approaches. Characterizing the mature neuronal environment is essential to understand the adult axonal guidance in order to complete the circuit reconstruction. To this end, we used mass spectrometry (MS)-based quantitative proteomics to characterize the proteomes of major nuclei of the adult visual system: suprachiasmatic nucleus (SCN), ventral and dorsal lateral geniculate nucleus (vLGN, dLGN) and superior colliculus (SCol)), as well as the optic chiasm. These analyses revealed the presence of guidance molecules and guidance-associated factors in the adult visual targets. Moreover, by performing bilateral optic nerve crush, we showed that the expression of specific proteins was significantly modulated by the injury in the visual targets, even in the ones most distal to the lesion site. On another hand, we found that the expression of guidance molecules was not modified upon injury and that regenerative axons are able to respond to guidance cues. Together, our results provide an extensive characterization of the molecular environment in intact and injured conditions.

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