Home

About

Advanced Search

Browse by Discipline

Scientific Societies

E-print Alerts

Add E-prints

E-print Network
FAQHELPSITE MAPCONTACT US


  Advanced Search  

 
coelom of stage 16/17 embryos by making a slit in the dorsal side of the embryo, between the prospective limb buds and lateral to the somites. The graft was inserted through the slit
 

Summary: coelom of stage 16/17 embryos by making a slit in the dorsal side of the embryo, between
the prospective limb buds and lateral to the somites. The graft was inserted through the slit
with the cut edge medial.
Recombinant limbs were prepared as described14
, except that mesenchyme was derived
from the distal-most 75100 mm of the anterior two-thirds of the wing bud and that
digestion with collagenase was omitted. Recombinant buds were grafted to the somites,
allowed to develop for 7 days, and stained with Victoria blue as described19
.
To rescue limb development in the absence of an AER, heparin acrylic beads were
incubated in 1 mg ml21
FGF4 (a gift from V. Rosen) at room temperature for 1 h, then
stored on ice until use. After removal of the AER, two heparin beads were attached to the
limb bud with platinum staples. One bead was placed at the posterior margin and the other
just anterior to the first. After attachment of beads, embryos were kept at room
temperature for 30 min before returning them to the incubator. Embryos were collected
after 6 days of incubation and fixed in 4% paraformaldehyde. Skeletons were stained with
0.02% alcian blue 8GX in 70% ethanol/30% acetic acid at 37 8C, then cleared in 0.5% KOH
and stored in glycerol.
Analysis of cell proliferation and death

  

Source: Abecasis, Goncalo - Department of Biostatistics, University of Michigan

 

Collections: Biology and Medicine; Mathematics