1. Whole-mount in situ Hybridization

2. Zebrafish Development

3. Function? Over-expressionknockdown DNA: using tissue specific promoter RNA: in vitro transcription Morpholino Dominant negative form Antagonist Whole mount in situ hybridization

5. Digoxigenin (DIG) DIG is a steroid hapten from Digitalis lanata ( 毛地黃 )

6. Structure of DIG-Modified Nucleotides Coupled via Alkali-Stable Ether Bond Formula: C 43 H 61 N 4 O 21 P 3 Li 4 Molecular Weight: 1090.7 DIG-UTP (R1=OH, R2=OH) DIG-dUTP (R1=OH, R2=H) DIG-ddUTP (R1= H, R2=H)

7. Non-Radioactive RNA-Labeling

9. TA vector SP6T7 Probe HindIII EcoRI HindIII

10. Detection methods Chemiluminescent detection  CDP-star or CSPD Colorimetric detection  NBT/BCIP Fluorescent detection

11. http://www.roche-applied- science.com/prodinfo_fst.htm?/DIG/dig_la beling03.htm http://zfin.org

12. mRNA: FGF8 (Fibroblast Growth Factor 8) Probe: Sef (Similar Expression of Fgf gene)

13. (Endocrine Reviews 2005, 26: 63-77)

14. (Cytokine & Growth Factor Reviews 2005, 16: 139-149)

15. (Nature Cell Biology 2002, 4: 170-174)

16. (Mechanisms of Development 2002, 119: 91-108)

17. (Development 2002, 129: 917-933)