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爪蟾不同生活周期示意图Xenopus laevis developmental stage chart

实验技术Protocol

下图概括爪蟾（Xenopus laevis）从受精卵到蝌蚪的关键发育阶段。示意图用于理解发育过程；精确 NF 分期应结合正文形态标准与 Xenbase 标准分期图谱。The illustration summarizes key stages from fertilized egg to tadpole. For precise NF staging, compare morphology with the Xenbase staging atlas.

受精卵、卵裂、囊胚、原肠胚、神经胚与蝌蚪的关键形态示意。
受精卵、卵裂、囊胚、原肠胚、神经胚与蝌蚪的关键形态示意。 科研示意图。Key developmental forms: egg, cleavage, blastula, gastrula, neurula and tadpole. Scientific schematic.
原图 1Original 1

NF 分期概要NF staging overview

NF 期NF stage发育阶段Developmental stage时间（25 ℃）Time at 25 ℃主要特征Key features
1–8受精卵 → 卵裂Fertilised egg → cleavage0–8 h第一、二次卵裂，8 细胞1st–2nd cleavage, 8-cell stage
8–9囊胚Blastula8–10 h囊腔形成，细胞数 ~1000Blastocoel forms, ~1000 cells
10–12原肠胚Gastrula10–14 h背唇（Spemann 组织者）出现，三胚层建立Dorsal lip (Spemann organizer) appears; three germ layers established
13–20神经胚Neurula14–22 h神经板、神经管闭合Neural plate forms, neural tube closes
21–35尾芽胚 → 游泳蝌蚪Tailbud → swimming tadpole1–3 d心跳、色素、尾芽伸展Heartbeat, pigmentation, tail extension
36–40自由游泳蝌蚪Free-swimming tadpole3–5 d消化道形成，开始摄食Gut forms, feeding begins

发育速度随温度显著变化：18–22 ℃ 较慢，25–27 ℃ 较快。实际分期须以形态为准，不能仅依据时间。Development rate varies significantly with temperature: slower at 18–22 ℃, faster at 25–27 ℃. Always stage by morphology, not time alone.

分期在实验中的应用Applications of staging

  • 显微注射时机：多选用 NF 1–2 期（单细胞或双细胞）进行 mRNA / Morpholino 注射，靶向胚胎全身；NF 8 期（8 细胞）可定向注射特定卵裂球，实现区域性标记。Injection timing: NF 1–2 (1- or 2-cell) for whole-embryo mRNA/Morpholino delivery; NF 8 (8-cell) for blastomere-targeted regional labelling.
  • 组织移植实验：NF 10.5 背唇是 Spemann 组织者实验的经典取材时期。Graft experiments: NF 10.5 dorsal lip is the classical stage for Spemann organizer assays.
  • 基因表达分析：ISH 和 IHC 通常覆盖 NF 10–40 各关键时间点，绘制基因时空表达图谱。Gene expression analysis: ISH and IHC typically span NF 10–40 key time points for spatiotemporal expression mapping.
  • 药物处理：将胚胎置于含目标化合物的盐溶液中，精确控制起始和终止发育时期。Drug treatment: expose embryos to compounds in saline solution with precise control of treatment window.

参考文献References

  1. Nieuwkoop PD, Faber J (1994) Normal Table of Xenopus laevis (Daudin), 2nd edn. Garland Publishing, New York. (Originally published 1956, North Holland Publishing Co., Amsterdam.)
  2. Harland R & Gerhart J (1997) Formation and function of Spemann's organizer. Annu Rev Cell Dev Biol 13:611–667.
  3. Sive HL, Grainger RM, Harland RM (2000) Early Development of Xenopus laevis: A Laboratory Manual. Cold Spring Harbor Laboratory Press.
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