Neural crest transplants
Tim Carl
Table of Methods
GFP-labelled crest transplant

Eggs are obtained from hormone-stimulated female Xenopus laevis and 
   fertilized in vitro following established lab procedures. 
Fertilized eggs are dejellied using 2% cysteine (pH 8.0) 
    and reared at room temperature in 10% Holtfreter's solution. 

Larvae are staged according to the normal table of Nieuwkoop & Faber (1967).

In Xenopus laevis, cranial neural crest migration begins in the late neurula (stage 19). 
   Tissue grafts are performed before migration, at stages 16 to early 19 
   (see Sadaghiani & Thiebaud, 1987). 

Embryos are placed in Petri dishes lined with 2% agar and 
  covered with 10% Holtfreter's + antibiotic (gentamycin, 50 µg/ml).

Small segments of the cranial neural folds from host embryos are ablated 
   and replaced by comparable segments from GFP-labeled donors 

Care must be taken to remove the overlaying ectoderm. 

Grafts is held in place initially with modeling clay and 
  the embryos are maintained at 16°C in Holtfreter's antibiotic. 

Specimens can be viewed using a stereo dissecting microscope equipped with epifluorescence optics. 

Table of Methods