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Maternal deposition of ApoA‐I‐mCherry transgene indicates an important role in early zebrafish development (1161.1)
Author(s) -
Reece Takisha,
Otis Jessica,
Farber Steven
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1161.1
Subject(s) - mcherry , transgene , biology , zebrafish , gene knockdown , apolipoprotein b , microbiology and biotechnology , genetically modified mouse , green fluorescent protein , genetics , gene , biochemistry , cholesterol
Our goal is to investigate the function of high‐density lipoprotein (HDL) particles of hepatic origin through in vivo visualization in the optically clear larval zebrafish ( Danio rerio ). We generated zebrafish stably expressing fluorescently tagged human apolipoprotein A‐I (ApoA‐I), a marker of HDL, through a liver‐specific promoter (LFABP:ApoA‐I‐mCherry; 6 lines). At 6‐days post fertilization (dpf), transgenic larvae show variable ApoA‐I‐mCherry localization: a subset of larvae show fluorescence in the liver, circulation, pronephric kidney and yolk syncytial layer, as expected; a second subset shows fluorescence in all of these regions except the liver. We hypothesized that variable ApoA‐I‐mCherry expression results from the first subset of larvae containing LFABP:ApoA‐I‐mCherry in their genome and that the second subset of larvae lack the transgene and are displaying maternally deposited ApoA‐I‐mCherry. Maternal RNA products deposited in embryos are degraded within hours, but the duration of maternal protein expression is less characterized. We pairwise crossed wild type and transgenic LFABP:ApoA‐I‐mCherry zebrafish and characterized ApoA‐I‐mCherry expression and localization. Only the progeny of LFABP:ApoA‐I‐mCherry females showed absence of ApoA‐I‐mCherry in the liver (n=12 experiments in 5 lines), supporting the conclusion that maternal deposition of ApoA‐I‐mCherry in non‐transgenic larvae created this phenotype. On‐going studies include Western Blot and PCR analyses to confirm and investigate the function of maternally deposited ApoA‐I in early development. Grant Funding Source : APS 2013 Frontiers in Physiology Research Teacher Fellowship