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Single cell transcriptomic and chromatin profiles suggest Layer Vb is the only layer with shared excitatory cell types in the medial and lateral entorhinal cortex

ABSTRACT: All brain functionality arises from the activity in neural circuits in different anatomical regions. These regions contain different circuits comprising unique cell types. An integral part to understanding neural circuits is a full census of the constituent parts, i.e., the neural cell types. This census can be based on different characteristics. Previously combinations of morphology and physiology, gene expression, and chromatin accessibility have been used in various cortical and subcortical regions. This has given an extensive yet incomplete overview of neural cell types. However, these techniques have not been applied to all brain regions. Here we apply single cell analysis of accessible chromatin on two similar but different cortical regions, the medial and the lateral entorhinal cortices. Even though these two regions are anatomically similar, their intrinsic and extrinsic connectivity are different. In 4136 cells we identify 20 different clusters representing different cell types. As expected, excitatory cells show regionally specific clusters, whereas inhibitory neurons are shared between regions. We find that several deep layer excitatory neuronal cell types as defined by chromatin profile are also shared between the two different regions. Integration with a larger scRNA-seq dataset maintains this shared characteristic for cells in Layer Vb. Interestingly, this Layer contains three clusters, two specific to either subregion and one shared between the two. These clusters can be putatively associated with particular functional and anatomical cell types found in this layer. This information is a step forwards into elucidating the cell types within the entorhinal circuit and by extension its functional underpinnings.

ORGANISM(S): Mus musculus

PROVIDER: GSE192567 | GEO | 2021/12/28

REPOSITORIES: GEO

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Project description:Studies of transcriptional networks in multi-cellular organisms usually focus on isolated cells and typically assume that the discovered gene networks represent those present in connected cells within a complex organ like the brain. However, similar cell types connected in diverse anatomical networks could differentially influence transcriptional networks. Here, we used laser capture microdissection, expression arrays, genome mapping, and computational inference to explore behaviorally regulated gene networks in the brains of awake, behaving songbirds producing a skilled motor behavior, singing. We found that at baseline, in the absence of singing, a large proportion of genes (17%, >3000) are differentially expressed in the different brain regions of the neural circuit that controls singing. These genes predominantly code for cell communication and connectivity proteins, and non-coding RNAs. Remarkably, the act of singing was associated with differential regulation of ~10% of the coding and non-coding genome. However, less than 1% of genes were singing-regulated in most brain regions and these were largely immediate early genes (IEGs), which peaked early, including the inducible transcription factors EGR1 and FOS. The remaining vast majority of behaviorally regulated gene expression was specific to one or a subset of brain regions, which peaked later. Promoters of the baseline, common, and diverse singing regulated gene clusters were enriched for different combinations of post-translationally activated transcription factors, like CREB, SRF, MEF2, MZF, and the IEG transcription factors. The results suggest that diverse cell-to-cell interactions and differential combinatorial binding of a small group of transcription factors may influence regional diversity of gene networks in seemingly similar cell types. Thus, in highly integrated neural circuits of intact behaving animals, transcriptional network diversity appears to be the rule, rather than the exception. Gene expression in Area X, HVC, LMAN, and Ra was measured before singing (0) or after singing for 0.5, 1, 2, 3, 4, 5, 6, and 7hours. Four-Six independent experiments were performed at each of the 9 timepoints.

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Single cell transcriptomic and chromatin profiles suggest Layer Vb is the only layer with shared excitatory cell types in the medial and lateral entorhinal cortex

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