Dataset Information

Deciphering the spatial-temporal molecular features of human developing hypothalamus at single-cell level

ABSTRACT: We revealed the molecular and cellular composition along spatial-temporal development of human hypothalamus by single-cell transcriptomic sequencing. The hypothalamus was manually micro-dissected from serial coronal 600-800μm thick slice using a vibratome. We excluded poor quality cells after the gene expression matrix was generated by Cell Ranger with Scanpy (V1.4.4) package. We identified 11 major transcriptomic types including neuroepithelium (NE), neural progenitors (NP), neurons, astrocytes, oligodendrocyte precursor cells (OPC), oligodendrocyte (OL), ependymocytes (Ependy), microglial cells, endothelial cells (Endoth), mural cells as well as vascular and leptomeningeal cells (VLMCs) that were characterzied with their respective classic markers. We uncovered molecular diversity of NP subtypes and constructed their developmental trajectory. We also identified various neuronal subpopulations mapped into distinct spatial location. Overall, our data provided cellular and molecular landscape of human hypothalamic development.

ORGANISM(S): Homo sapiens

PROVIDER: GSE169109 | GEO | 2021/12/07

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Purpose: We aimed to reconstruct the development formation of cellular identities' heterogeneity in the hypothalamus with single-cell RNA sequencing. We planned to track branching process of neurogenesis and follow the formation of cell identities. Results: We integrate single-cell RNA-seq data on 51,199 cells of ectodermal origin, representing critical stages of pre- and postnatal hypothalamus development, supported by anatomical lineage reconstruction to show that bulk neurogenesis in the hypothalamus, dominated by Notch signalling and its interaction with Ascl1, persists until the neonatal period to produce molecularly distinct astrocytes, ependyma, tanycytes and 30 protoclusters of neurons. Alike in adults, the assignment of neurotransmitters and neuropeptides together with transcription factors as positional marks for each neuronal cluster is sufficient to establish the spatially-resolved taxonomy of hypothalamic neurons. By deducing gene regulatory networks, exemplified by hierarchical gene clusters driven by Onecut2/Onecut3 transcription factors for dopamine neurons, we resolve how neuronal lineages diverge into functionally non-overlapping endocrine subtypes. Dopamine neurons are used to illustrate molecular convergence with their shared neurotransmitter identity representing a mismatch to their molecular distances on multidimensional vector spaces. We demonstrate the existence of non-canonical chemotropic axes in the hypothalamus, particularly sequential ligand-receptor switching in the unexpectedly abundant Slit/Robo cassette. Slit2 is identified in ventricular ependymocytes and implicated in repulsing new-born neurons that sequentially express Robo1 and Robo2 receptors for migration and differentiation. Conclusions: Overall, our unbiased map of hypothalamic cell types and extracellular guidance cues uncovers molecular principles shaping the developmental architecture of the mammalian hypothalamus and rationalise how hypothalamic neuronal heterogeneity is transformed into a multimodal neural unit that retains a virtually infinite adaptive potential throughout life.

Project description:The ependyma lining the third ventricle (3V) in the mediobasal hypothalamus is recognized as a critical player in controlling energy balance and glucose homeostasis. Its molecularly distinct cell types, namely diverse tanycyte subpopulations and typical ependymal cells, confer a high functional heterogeneity and plasticity to this complex ependymal system, but the molecular mechanisms governing its features, especially in the context of metabolic regulation, are still not fully understood.Here, 5481 hypothalamic ependymocytes were cataloged using FACS-assisted single-cell RNA sequencing from fed, 12h-fasted, and 24h-fasted adult male mice. We first observed the limitations of using standard clustering analysis to characterize the different ependymal cell subpopulations lining the 3V. Indeed, while typical ependymal cells and β2-tanycytes are sharply defined at the molecular level using this approach, other subpopulations (i.e., β1- and α-tanycytes) display fuzzy boundaries and very few specific markers. Moreover, 12h- and 24h-fasting dynamically modulate gene expression, increasing tanycyte subgroup heterogeneity. To better analyze the dynamic changes in tanycytes’ gene expression profile, we implemented a pseudospatial analysis based on 3V neuroanatomical distribution, which we validated using in-situ hybridization, immunohistochemistry, and histologic staining. This approach improved the characterization of tanycyte markers, refining our knowledge of specific versus shared features and the segregation of subgroup-specific versus general tanycyte functions. Importantly, we showed that fasting dynamically shifts patterns in gene expression and transcription activity along the 3V, inducing metabolic and functional switches in tanycyte subpopulations, particularly regarding lipid metabolism. Additionally, numerous functions related to tanycyte-neuron and tanycyte-synapse interactions, notably through Nrxn1 expression, present a functional dorso→ventral shift towards the arcuate nucleus during fasting. Altogether, this data shows that changes in energy status lead to a spatial redistribution of distinct cell type-specific responses along the 3V and gives new insights into molecular and functional diversity and plasticity within the tanycyte population.

Dataset Information

Deciphering the spatial-temporal molecular features of human developing hypothalamus at single-cell level

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets