Effects of epidermal growth factor (EGF), transforming growth factor-α (TGFα), and 2,3,7,8- tetrachlorodibenzo-p-dioxin on fusion of embryonic palates in serum-free organ culture using wild-type, EGF knockout, and TGFα knockout mouse strains
Effects of epidermal growth factor (EGF), transforming growth factor-α (TGFα), and 2,3,7,8- tetrachlorodibenzo-p-dioxin on fusion of embryonic palates in serum-free organ culture using wild-type, EGF knockout, and TGFα knockout mouse strains
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DOI：
10.1002/bdra.20133
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发表时间：
2005-06
影响因子：
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通讯作者：
B. Abbott;A. Buckalew;Korin E. Leffler
中科院分区：
文献类型：
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作者：
B. Abbott;A. Buckalew;Korin E. Leffler
背景 2,3,7,8-四氯二苯并-对二恶英 (TCDD) 对小鼠具有致畸作用，产生腭裂 (CP)。 TCDD 暴露会破坏上腭中表皮生长因子 (EGF) 受体、EGF 和转化生长因子-α (TGFα) 的表达，并影响内侧上皮细胞的增殖和分化。 EGF 敲除胚胎对 TCDD 诱导 CP 的敏感性较低。这项研究使用上颚器官培养来检验 EGF 能够对 TCDD 做出反应的假设。方法 将来自野生型 (WT)、EGF 敲除、C57BL/6J 和 TGFα 敲除胚胎的中面组织在妊娠第 12 天 (GD) 置于器官培养中。腭外植体在无血清 Bigger's (BGJ) 培养基中培养 4 天，其中含有 0.1% 二甲基亚砜 (DMSO) 或 1 × 10−8 M TCDD（含或不含 2 ng） EGF/ml，1 或 2 ng TGFα/ml。在培养第 4 天评估对腭融合的影响。使用 Luminex 技术测定外植体和培养基中的 EGF 水平。结果 在无血清对照培养基中，所有菌株的味觉融合。用 TCDD（无 EGF）培养的 EGF 敲除腭融合，但用 TCDD + 2 ng EGF/ml 培养的那些未能融合（与对照或不含 EGF 的 TCDD 相比，p < 0.05）。当与 TCDD + 2 ng TGFα/ml 一起培养时，TGFα 敲除的腭未能融合。培养 24 小时后，组织中的 EGF 水平增加并在培养基中积累。结论 这项研究表明，向 EGF 敲除小鼠的上颚提供 EGF 可以恢复对 TCDD 的反应。这些研究支持这样的假设：TCDD 诱导 CP 的机制是通过 EGFR 途径介导的。出生缺陷研究（A 部分），2005 年。2005 年出版 Wiley-Liss, Inc.
BACKGROUND 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is teratogenic in mice, producing cleft palate (CP). TCDD exposure disrupts expression of epidermal growth factor (EGF) receptor, EGF, and transforming growth factor-α (TGFα) in the palate and affects proliferation and differentiation of medial epithelial cells. EGF knockout embryos are less susceptible to the induction of CP by TCDD. This study used palate organ culture to examine the hypothesis that EGF enables a response to TCDD. METHODS The midfacial tissues from wild-type (WT), EGF knockout, C57BL/6J, and TGFα knockout embryos were placed in organ culture on gestational day (GD) 12. Palatal explants were cultured for 4 days in serum-free Bigger's (BGJ) medium with 0.1% dimethyl sulfoxide (DMSO) or 1 × 10−8 M TCDD with or without 2 ng of EGF/ml, 1 or 2 ng of TGFα/ml. Effects on palatal fusion were evaluated on day 4 of culture. EGF levels in explants and medium were determined using Luminex technology. RESULTS In serum-free, control medium, palates from all of the strains fused. EGF knockout palates cultured with TCDD (no EGF) fused, but those cultured with TCDD + 2 ng of EGF/ml failed to fuse (p < 0.05 vs. control or TCDD without EGF). TGFα knockout palates failed to fuse when cultured with TCDD + 2 ng of TGFα/ml. EGF levels increased in tissue and accumulated in the medium after 24 hr of culture. CONCLUSIONS This study demonstrated that providing EGF to the palates of EGF knockout mice restored the response to TCDD. These studies support the hypothesis that the mechanism for induction of CP by TCDD is mediated via the EGFR pathway. Birth Defects Research (Part A), 2005. Published 2005 Wiley-Liss, Inc.