Characterization of a new, inducible transgenic mouse model with GFP expression in melanocytes and their precursors
| Autor: | Maira Castaneda, Shunlin Jiang, Ganesh Diwakar, Ronna Hertzano, Bishal Tandukar, Sandeep S. Joshi, Thomas J. Hornyak |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine Genetically modified mouse Mice Transgenic Biology Melanocyte Stem cell marker Article Green fluorescent protein Mice 03 medical and health sciences Melanocyte differentiation Genetics medicine Animals Cell Lineage Promoter Regions Genetic Molecular Biology Embryonic Stem Cells Protein Synthesis Inhibitors Membrane Glycoproteins Pigmentation Gene Expression Regulation Developmental Tetracycline Cell sorting Flow Cytometry Hair follicle Cell biology Mice Inbred C57BL body regions 030104 developmental biology medicine.anatomical_structure Lac Operon Neural Crest Models Animal Melanocytes Female Stem cell Oxidoreductases Hair Follicle gp100 Melanoma Antigen Developmental Biology |
| Zdroj: | Gene Expression Patterns. 27:76-84 |
| ISSN: | 1567-133X |
| DOI: | 10.1016/j.gep.2017.10.003 |
| Popis: | Melanocytes are neural crest-derived cells that are responsible for mammalian hair follicle (HF) pigmentation. The Dct-LacZ transgenic mouse is extensively used to study melanocyte biology but lacks conditionally-inducible labelling and fluorescent labelling, enabling specific, viable isolation of melanocytes using fluorescence-activated cell sorting (FACS). Here, we have generated a Tet-off bitransgenic mouse model, Dct-H2BGFP, containing Dct-tTA and TRE-H2BGFP transgenes. Characterization of Dct-H2BGFP mice confirmed a pattern of Dct-H2BGFP expression in melanoblasts, melanocyte stem cells (McSCs), and terminally differentiated melanocytes similar to the expression pattern of previously published mouse models Dct-LacZ and iDct-GFP. GFP expression is regulated by doxycycline. GFP is shown to co-localize with melanocyte label-retaining cells (LRCs) identified through BrdU retention. The GFP-expressing cells identified in vivo in the bulge and the secondary hair germ of telogen HFs of Dct-H2BGFP mice express the melanocyte and melanocyte stem cell markers Dct and Kit. Using Dct-H2BGFP mice, we separated GFP-expressing cells from the telogen HF based on FACS and showed that GFP-expressing cells express high levels of Kit and Dct, and lower levels of HF epithelial keratin genes. We also show that GFP-expressing cells express high levels of the melanocyte differentiation genes Tyr, Tyrp1, and Pmel17, further substantiating their identity within the melanocyte lineage. Thus, Dct-H2BGFP mice are not only useful for the in vivo identification of melanocytic cells, but also for isolating them viably and studying their molecular and biological properties. |
| Databáze: | OpenAIRE |
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