A doxycycline-inducible, tissue-specific aromatase-expressing transgenic mouse
Autor: | Wah Chin Boon, Margaret M. Bills, Jenny D. Y. Chow, John T. Price, Evan R. Simpson |
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Rok vydání: | 2011 |
Předmět: |
Male
Genetically modified mouse DNA Complementary Microinjections medicine.drug_class Transgene Genetic Vectors Mice Transgenic Gene Expression Regulation Enzymologic Salivary Glands Mice Aromatase Luciferases Firefly Cell Line Tumor Gene expression Genetics medicine Animals Humans Bioluminescence imaging Luciferase Transgenes Cloning Molecular Mammary Glands Human Promoter Regions Genetic Enzyme Assays Doxycycline biology Reverse Transcriptase Polymerase Chain Reaction Seminal Vesicles Molecular biology Enzyme Activation Estrogen Luminescent Measurements biology.protein Female Animal Science and Zoology Agronomy and Crop Science Plasmids Biotechnology medicine.drug |
Zdroj: | Transgenic Research. 21:415-428 |
ISSN: | 1573-9368 0962-8819 |
Popis: | Aromatase converts androgens to estrogens and it is expressed in gonads and non-reproductive tissues (e.g. brain and adipose tissues). As circulating levels of estrogens in males are low, we hypothesize that local estrogen production is important for the regulation of physiological functions (e.g. metabolism) and pathological development (e.g. breast and prostate cancers) by acting in a paracrine and/or intracrine manner. We generated a tissue-specific doxycycline-inducible, aromatase transgenic mouse to test this hypothesis. The transgene construct (pTetOAROM) consists of a full-length human aromatase cDNA (hAROM) and a luciferase gene under the control of a bi-directional tetracycline-responsive promoter (pTetO), which is regulated by transactivators (rtTA or tTA) and doxycycline. Our in vitro studies using MBA-MB-231tet cells stably expressing rtTA, showed that doxycycline treatment induced transgene expression of hAROM transcripts by 17-fold (P = 0.01), aromatase activity by 26-fold, (P = 0.0008) and luciferase activity by 9.6-fold (P = 0.0006). Pronuclear microinjection of the transgene generated four pTetOAROM founder mice. A male founder was bred with a female mammary gland-specific rtTA mouse (MMTVrtTA) to produce MMTVrtTA-pTetOAROM double-transgenic mice. Upon doxycycline treatment via drinking water, human aromatase expression was detected by RT-PCR, specifically in mammary glands, salivary glands and seminal vesicles of double-stransgenic mice. Luciferase expression and activity was detected in these tissues by in vivo bioluminescence imaging, in vitro luciferase assay and RT-PCR. In summary, we generated a transgenic mouse model that expresses the human aromatase transgene in a temporal- and spatial-specific manner, which will be a useful model to study the physiological importance of local estrogen production. |
Databáze: | OpenAIRE |
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