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Published on: October 10, 2011
Involvement of EAT/mcl-1, an anti-apoptotic bcl-2-related gene, in murine embryogenesis and human development
1Department of Pathology, Department of Obstetrics and Gynecology, Laboratory Animal Center, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan.
Abstract:
Apoptosis plays an important regulatory role in mammalian embryogenesis and development. EAT/mcl-1 (EAT), an anti-apoptotic bcl-2-related gene, was isolated during the early differentiation of a human embryonal carcinoma cell line, an event which serves as a model of early embryogenesis. EAT is involved in apoptotic regulation and is believed to also function as an immediate-early gene. Thus it was hypothesized that EAT would be expressed during early embryogenesis and would be involved in the regulation of apoptosis during this critical period. To clarify this early expression, two antibodies to EAT were generated by use of immunizing oligopeptide (aa 37-55) and recombinant protein (aa 31-229) for use in immunohistochemistry and immunoblotting, respectively. With these antibodies, we then determined EAT expression during murine embryogenesis and in human development, using human fetal tissue of 6 to 23 gestational weeks. During murine embryogenesis, the EAT protein was found to be rapidly induced after fertilization, to peak at the 2-cell stage, to remain constant until the 8-cell stage, and then to decrease to below unfertilized egg levels in blastocysts. EAT expression patterns in early human development were found to essentially overlap those observed in adult tissues which suggest that EAT expression continues until adulthood in terminally differentiated tissues. Among tissues distinct to fetal development, EAT was detected in the mesonephric (Wolffian) duct and paramesonephric (Müllerian) duct. It is also noteworthy that prominent EAT immunoreactivity was also observed in large primary oocytes in 21-week fetal ovary, but was not detected in primordial germ cells in 23-week fetal testis. In summary, EAT expression was detected in hematopoietic, epithelial, neural, endocrine, and urogenital cells; this provides evidence that EAT, as an anti-apoptotic molecule, possibly functions to regulate apoptosis during development in these systems.
Insights
The anti-apoptotic gene EAT/mcl-1 (EAT) is expressed during early mammalian embryogenesis, playing a key role in regulating apoptosis. Its expression patterns suggest a vital function in embryonic development and tissue differentiation.
Area of Science:
- Developmental Biology
- Molecular Biology
- Cell Biology
Background:
- Apoptosis is crucial for mammalian embryogenesis.
- EAT/mcl-1 (EAT) is an anti-apoptotic gene identified during early human embryonal carcinoma cell differentiation.
- EAT is hypothesized to be an immediate-early gene involved in regulating apoptosis during development.
Purpose of the Study:
- To investigate the expression of EAT during early murine and human development.
- To determine if EAT is involved in the regulation of apoptosis in embryonic tissues.
Main Methods:
- Generated two antibodies against EAT for immunohistochemistry and immunoblotting.
- Analyzed EAT protein expression in murine embryos post-fertilization through blastocyst stages.
- Examined EAT expression in human fetal tissues from 6 to 23 gestational weeks.
Main Results:
- EAT protein rapidly induced after fertilization in mice, peaking at the 2-cell stage and decreasing by the blastocyst stage.
- EAT expression in human development largely mirrored adult tissue patterns, suggesting continued expression.
- EAT detected in fetal urogenital ducts (mesonephric and paramesonephric), fetal ovary oocytes, but not fetal testis germ cells.
Conclusions:
- EAT is expressed during early mammalian embryogenesis and human development.
- EAT protein is present in various fetal cell types, including hematopoietic, epithelial, neural, endocrine, and urogenital cells.
- EAT's role as an anti-apoptotic molecule suggests it regulates apoptosis in developing mammalian systems.
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