Calcium dynamics and chromatin remodelling underlie heterogeneity in prolactin transcription
| Autor: | Claire V. Harper, Jayne C Charnock, Anne V McNamara, David G. Spiller, Julian R. E. Davis, Michael R. H. White |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Cell type prolactin Transcription Genetic Population chemistry.chemical_element 030209 endocrinology & metabolism Calcium Cell Line Histones 03 medical and health sciences Genetic Heterogeneity 0302 clinical medicine Endocrinology Animals education Molecular Biology education.field_of_study calcium biology Research Acetylation Cell sorting Chromatin Assembly and Disassembly Prolactin Cell biology Chromatin Rats single cell 030104 developmental biology Histone chemistry biology.protein chromatin Single-Cell Analysis heterogeneity transcription Chromatin immunoprecipitation |
| Zdroj: | Journal of Molecular Endocrinology |
| ISSN: | 1479-6813 |
| Popis: | Pituitary cells have been reported to show spontaneous calcium oscillations and dynamic transcription cycles. To study both processes in the same living cell in real time, we used rat pituitary GH3 cells stably expressing human prolactin-luciferase or prolactin-EGFP reporter gene constructs loaded with a fluorescent calcium indicator and measured activity using single-cell time-lapse microscopy. We observed heterogeneity between clonal cells in the calcium activity and prolactin transcription in unstimulated conditions. There was a significant correlation between cells displaying spontaneous calcium spikes and cells showing spontaneous bursts in prolactin expression. Notably, cells showing no basal calcium activity showed low prolactin expression but elicited a significantly greater transcriptional response to BayK8644 compared to cells showing basal calcium activity. This suggested the presence of two subsets of cells within the population at any one time. Fluorescence-activated cell sorting was used to sort cells into two populations based on the expression level of prolactin-EGFP however, the bimodal pattern of expression was restored within 26 h. Chromatin immunoprecipitation showed that these sorted populations were distinct due to the extent of histone acetylation. We suggest that maintenance of a heterogeneous bimodal population is a fundamental characteristic of this cell type and that calcium activation and histone acetylation, at least in part, drive prolactin transcriptional competence. |
| Databáze: | OpenAIRE |
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