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Single-cell transcriptomics of neonatal mouse anterior pituitary in steady-state conditions and after transgenically inflicted local damage

ABSTRACT: The mouse pituitary gland undergoes vivid maturation immediately after birth. During this process, the local stem cell compartment shows signs of activation including elevated abundance and expression of stemness pathways when compared to adult pituitary stem cells. In addition, we found that the neonatal pituitary displays high regeneration efficiency, as occurring following diphtheria toxin (DT)-triggered endocrine cell-ablation injury using the GHCre/iDTR mouse model (expressing Cre recombinase under control of the growth hormone (Gh) promoter, as well as Cre-inducible (floxed) DT receptor (iDTR)). This regeneration is more pronounced than in older mice, which is in line with the activated (stem cell) nature of the neonatal gland. However, it is not known what molecular mechanisms underlie the stem cell activation status in the neonatal gland. Here, we set out to decode the stem cells’ phenotype during the neonatal pituitary maturation stage starting from single cell RNA-sequencing (scRNA-seq) interrogations of neonatal (postnatal day 7, PD7) normal (undamaged) and damaged pituitary (more specifically, its major endocrine anterior lobe).

INSTRUMENT(S): Illumina NovaSeq 6000

ORGANISM(S): Mus musculus

SUBMITTER: Florian Hermans

PROVIDER: E-MTAB-11337 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Single-cell transcriptional profiling of young-adult and aging (middle-aged) mouse anterior pituitary in steady-state conditions and after transgenically inflicted local damage

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Project description:Transcription factors and signaling pathways that regulate stem cells and specialized hormone-producing cells in the pituitary gland have been the subject of intense study and have yielded a mechanistic understanding of pituitary organogenesis and disease. Yet, the regulation of stem cell proliferation and differentiation, the heterogeneity among specialized hormone-producing cells, and the role of non-endocrine cells in the gland remain important, unanswered questions. Recent advances in single-cell RNA sequencing (scRNAseq) technologies provide new avenues to address these questions. We performed scRNAseq on approximately 13,663 cells pooled from six whole pituitary glands of 7-week-old C57BL/6 male mice. We identified pituitary endocrine and stem cells in silico, as well as other support cell-types such as endothelia, connective tissue, and red and white blood cells. Differential gene expression analyses identify known and novel markers of pituitary endocrine and stem cell populations. We demonstrate the value of scRNAseq by in vivo validation of a novel gonadotrope-enriched marker, Foxp2. We present novel scRNAseq data of in vivo pituitary tissue, including data from agnostic clustering algorithms which suggest the presence of a somatotrope subpopulation enriched in sterol/cholesterol synthesis genes. At the same time, we show that incomplete transcriptome annotation can cause false negatives on some scRNAseq platforms that only generate 3’ transcript end sequences, and use in vivo data to recover reads of the pituitary transcription factor Prop1. Ultimately, scRNAseq technologies represent a significant opportunity to address longstanding questions regarding the development and function of the different populations of the pituitary gland throughout life.

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