Chronological Analysis With Fluorescent Timer Reveals Unique Features of Newly Generated β-Cells
| Autor: | Takeshi Miyatsuka, Taka-aki Matsuoka, Shugo Sasaki, Michael S. German, Fumiyo Kubo, Iichiro Shimomura, Hirotaka Watada, Manami Hara |
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| Rok vydání: | 2014 |
| Předmět: |
Endocrinology
Diabetes and Metabolism Cellular differentiation Mice Transgenic Nerve Tissue Proteins Biology Neogenesis Flow cytometry Cell therapy Transcriptome Mice Insulin-Secreting Cells Basic Helix-Loop-Helix Transcription Factors Internal Medicine medicine Animals Insulin Paired Box Transcription Factors Progenitor cell Promoter Regions Genetic Progenitor Homeodomain Proteins Genetics medicine.diagnostic_test Cell Differentiation SOX9 Transcription Factor Cell cycle Flow Cytometry biology.organism_classification Cell biology Islet Studies |
| Zdroj: | Diabetes |
| ISSN: | 1939-327X 0012-1797 |
| DOI: | 10.2337/db13-1312 |
| Popis: | Although numerous studies have uncovered the molecular mechanisms regulating pancreas development, it remains to be clarified how β-cells arise from progenitors and how recently specified β-cells are different from preexisting β-cells. To address these questions, we developed a mouse model in which the insulin 1 promoter drives DsRed-E5 Timer fluorescence that shifts its spectrum over time. In transgenic embryos, green fluorescent β-cells were readily detected by FACS and could be distinguished from mature β-cells only until postnatal day 0, suggesting that β-cell neogenesis occurs exclusively during embryogenesis. Transcriptome analysis with green fluorescent cells sorted by FACS demonstrated that newly differentiated β-cells highly expressed progenitor markers, such as Sox9, Neurog3, and Pax4, showing the progenitor-like features of newborn β-cells. Flow cytometric analysis of cell cycle dynamics showed that green fluorescent cells were mostly quiescent, and differentiated β-cells were mitotically active. Thus, the precise temporal resolution of this model enables us to dissect the unique features of newly specified insulin-producing cells, which could enhance our understanding of β-cell neogenesis for future cell therapy. |
| Databáze: | OpenAIRE |
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