Single-cell molecular and cellular architecture of neurohypophyseal cell types in adult mouse
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ABSTRACT: The neurohypophysis (NH), located at the posterior lobe of the pituitary, is a major neuroendocrine tissue, which mediates osmotic balance, blood pressure, reproduction, and lactation by means of releasing the neurohormones oxytocin and arginine-vasopressin from the brain into the peripheral blood circulation. The major cellular components of the NH are hypothalamic axonal termini, fenestrated endothelia and pituicytes, the resident astroglia. However, despite the physiological importance of the NH, the exact molecular signature defining neurohypophyseal cell types and in particular the pituicytes, remains unclear. Using single cell RNA sequencing, we captured seven distinct cell types in the NH and intermediate lobe (IL) of adult male mouse. We revealed novel pituicyte markers showing higher specificity than previously reported markers. Multiplexing single molecule in situ hybridization revealed spatial organization of the major cell types implying intercellular communications. We present a comprehensive molecular and cellular characterization of neurohypophyseal cell-types serving as a valuable resource for further functional research.
Project description:We have used microarrays to comprehensively describe the transcriptomes of the supraoptic nucleus (SON), the paraventricular nucleus (PVN) and the neurointermediate lobe (NIL) of adult male Sprague-Dawley (SD) and Wistar-Kyoto (WKY) rats, as well as the paraventricular nucleus of Wistar (WIST) rats. Comparison of these gene lists has enabled us to identify surprisingly large differences in hypothalamo-neurohypophyseal system gene expression patterns in these three strains. We have also shown that different transcript populations are enriched in the PVN and the SON of SD and WKY rats. The transcriptome differences catalogued here may be molecular substrates for the neuro-humoral phenotypic differences exhibited by different strains of rats. Keywords: Transcriptome, Hypothalamo-neurohypophyseal system, Genetic
Project description:The pituitary is the master neuroendocrine tissue, which regulates body homeostasis. It consists of the adenohypophysis (AH), which harbors hormones producing cells and the neurohypophysis (NH), which relays the direct passage of hormones from the brain to the periphery. We previously characterized the mouse NH cell identities (Chen Q et al., 2020). However, the zebrafish NH cell types remained unknown owning to surgical inaccessible and limited amount of the tissue. Using Mars-Seq single-cell sequencing (Jaitin et al., 2014), we characterized the cell types of the neurohypophyseal enriched population from the juvenile and adult zebrafish pituitaries, which laid the foundation for further lineage and functional research in this study.
Project description:The hypothalamo-neurohypophyseal system (HNS) is an interface through which the brain regulates body homeostasis by means of releasing the hypothalamic neurohormones oxytocin and arginine-vasopressin to the general circulation. The basic components of the HNS are the hypothalamic axonal projections, endothelial blood vessels and astroglial-like cells, termed pituicytes. These three tissue types converge and interact at the ventral forebrain to establish an efficient neuro-vascular interface, which allows the release of neurohormones from the brain to the periphery. However, the molecular blueprint of pituicytes-specific genes is still unknown. We have labelled and isolated zebrafish pituicytes cells and have identified their molecular signature.
Project description:We have used microarrays to comprehensively describe the transcriptomes of the supraoptic nucleus (SON), the paraventricular nucleus (PVN) and the neurointermediate lobe (NIL) of adult male Sprague-Dawley (SD) and Wistar-Kyoto (WKY) rats, as well as the paraventricular nucleus of Wistar (WIST) rats. Comparison of these gene lists has enabled us to identify surprisingly large differences in hypothalamo-neurohypophyseal system gene expression patterns in these three strains. We have also shown that different transcript populations are enriched in the PVN and the SON of SD and WKY rats. The transcriptome differences catalogued here may be molecular substrates for the neuro-humoral phenotypic differences exhibited by different strains of rats. Experiment Overall Design: For each experimental group (SON-WKY, PVN-WKY, NIL-WKY, SON-SD, PVN-SD, NIL-SD, PVN-WISTAR) five chips were hybridised with independantly pooled RNA from a biological n=5.
Project description:This SuperSeries is composed of the following subset Series:; GSE3110: Comprehensive description of the transcriptome of hypothalamo-neurohypophyseal system in euhydrated and dehydrated rat; GSE3111: Comprehensive description of the transcriptome of PVN in euhydrated and dehydrated rat; GSE3125: Comprehensive description of the transcriptome of neurointermediate lobe in euhydrated and dehydrated rat Experiment Overall Design: Refer to individual Series
Project description:The “ship of the desert”, the one-humped Arabian camel (Camelus dromedarius), has a remarkable capacity to survive in conditions of extreme heat without needing to drink water. One of the ways that this is achieved is through the actions of the antidiuretic hormone vasopressin (AVP) and the natriuretic hormone oxytocin (OXT), both of which are made in a specialised part of the brain called the hypothalamo-neurohypophyseal system (HNS), but exert their effects at the level of the kidney to, respectively, provoke water conservation and salt excretion. Interestingly, our electron microscopy studies have shown that the ultrastructure of the camel HNS changes according to season, suggesting that in the arid conditions of summer the dromedary’s HNS is in a state of permanent activation, in preparation for the likely prospect of water deprivation. Based on our camel genome sequence, we have carried out an RNAseq analysis of the camel HNS in summer and winter.
Project description:The mammalian pituitary gland is a complex organ consisting of hormone-producing cells, anterior lobe folliculostellate cells (FSCs), posterior lobe pituicytes, vascular pericytes and endothelial cells, and Sox2-expressing stem cells. We present single-cell RNA sequencing and immunohistofluorescence analyses of pituitary cells of adult female rats with a focus on the transcriptomic profiles of nonhormonal cell types. Samples obtained from whole pituitaries and separated anterior and posterior lobe cells contained all expected pituitary resident cell types and lobe-specific vascular cell subpopulations. FSCs and pituicytes expressed S100B, ALDOC, EAAT1, ALDH1A1, and VIM genes and proteins, as well as other astroglial marker genes, some common and some cell type-specific. We also found that the SOX2 gene and protein were expressed in ~15% of pituitary cells, including FSCs, pituicytes, and a fraction of hormone-producing cells, arguing against its stem cell specificity. FSCs comprised two Sox2-expressing subclusters; FS1 contained more cells but lower genetic diversity, while FS2 contained proliferative cells, shared genes with hormone-producing cells, and expressed genes consistent with stem cell niche formation, regulation of cell proliferation and stem cell pluripotency, including the Hippo and Wnt pathways. FS1 cells were randomly distributed in the anterior and intermediate lobes, while FS2 cells were localized exclusively in the marginal zone between the anterior and intermediate lobes. These data indicate the identity of the FSCs as anterior pituitary-specific astroglia, with FS1 cells representing differentiated cells equipped for classical FSC roles and FS2 cells exhibiting additional stem cell-like features.