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The role of the alpha‐tocopherol transfer protein during zebrafish embryogenesis
Author(s) -
Miller Galen William,
Labut Edwin M,
Lebold Katie M,
Barton Carrie,
Tanguay Robert L,
Traber Maret G
Publication year - 2011
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.25.1_supplement.937.3
Subject(s) - zebrafish , morpholino , gene knockdown , messenger rna , embryo , microbiology and biotechnology , yolk sac , alternative splicing , biology , embryogenesis , andrology , chemistry , genetics , cell culture , gene , medicine
The alpha‐tocopherol transfer protein (TTP) is required for vitamin E bioavailability and transfer to the blood stream. The finding that TTP is expressed in mammalian uterine and placental cells emphasizes the significance of TTP during embryogenesis. To study TTP's role in fetal development, a non‐placental model is essential. The zebrafish meets these requirements. Expression of TTP mRNA in zebrafish embryos increases 10‐fold within the first 10 h of zebrafish development and increases again between 24 and 36 h post fertilization (hpf). TTP was knocked down using morpholinos (MO) designed to inhibit TTP mRNA processing by interfering with proper splicing or translational initiation. Attenuating TTP by blocking proper mRNA splicing in the zebrafish embryos caused a distinct “pinched” phenotype in >50% of the embryos, observed at 8 hpf, resulting from an irregular yolk sac engulfment, suggesting membrane instability. Corresponding with the second increase in TTP mRNA, both MOs at 24 hpf caused severe cell death in the developing brain (>50% and >95%, splice‐blocking and translation inhibition respectively) while concentration matched controls caused less (<5%). To control for non‐specific effects we injected a MO against p53, which resulted in less mortality by 8 hpf while preserving the TTP knockdown effects. Thus, TTP is vital for embryonic survival. Funding: NICHD 5R01HD062109‐02