Robustness in the feedback control of the retinoic acid network response to environmental disturbances (BioMark-qRT-PCR). Robustness in the feedback control of the retinoic acid network response to environmental disturbances (BioMark-qRT-PCR)

Retinoic acid (RA) is a central developmental signal whose perturbation results in teratogenic outcomes. RA is tightly regulated during early embryonic development to achieve fine-tuning of the instructive and permissive signals regulating multiple developmental processes including head formation. We aim to identify the key mechanisms underlying the homeostatic regulation and tuning of the retinoic acid signaling levels, the disruption of which in the head lead to wide ranging developmental malformations including microcephaly and other craniofacial defects. We performed transient physiological RA signaling disturbances during Xenopus laevis embryogenesis followed by kinetic gene expression analysis of the recovery at multiple time points. Overall design: We considered multiple experimental perturbations including exogenous addition of 10 nM RA, inhibition of RA production by 50 uM DEAB (an inhibitor of RALDH2 enzyme) and no exposure control. We obtained data from six independent clutches for each experimental condition. The embryos were isolated at multiple time points (t=0, 1.5, 3, and 4.5 hours) following two hours of perturbation and a wash.

视黄酸（Retinoic acid, RA）是核心发育信号分子，其扰动可引发致畸后果。在早期胚胎发育过程中，RA受到严格调控，以精准微调指导性与允许性信号，这些信号调控包括头部形成在内的多种发育进程。本研究旨在阐明视黄酸信号水平的稳态调控与微调机制——该信号在头部的紊乱会导致广泛的发育畸形，包括小头症及其他颅面缺陷。我们在非洲爪蟾（Xenopus laevis）胚胎发生过程中施加瞬时生理性RA信号扰动，随后在多个时间点对胚胎的恢复过程进行动力学基因表达分析。

总体实验设计：我们设置了多种实验扰动条件，包括外源施加10 nM视黄酸、通过50 μM DEAB（一种RALDH2酶抑制剂）抑制RA生成，以及未暴露的空白对照组。每种实验条件均取自6个独立卵群。在施加扰动并清洗2小时后，我们在多个时间点（t=0、1.5、3和4.5小时）分离胚胎样本。