Producing exposed coat-free embryos.
Production of embryos that are free of tough outer coats facilitates studies that are not possible with embryos surrounded by impenetrable envelopes. This report describes a new procedure for preventing formation of fertilisation membranes in the sea urchin (Lytechinus pictus) model. This procedure involves treating unfertilised eggs with the enzyme alpha-amylase, which cleaves alpha-1,4 glucosidic bonds in the vitelline layer. A major advantage of this method is that it is very well defined and completely controllable with alpha-amylase inhibitor. The results suggest that intact alpha-1,4 glucosidic bonds are essential for vitelline layer integrity required for formation of the fertilisation membrane. Eggs treated with alpha-amylase possessed the same surface lectin receptors as untreated eggs and, as shown by light and transmission electron microscopy, produced healthy, cleaving embryos that were free of fertilisation envelopes.
Daily
M F
MF
Department of Biology, California State University, Northridge 91330-8303, USA.
Latham
V H
VH
Garcia
C M
CM
Hockman
C L
CL
Chun
H
H
Oppenheimer
M L
ML
West
S P
SP
Rostamiany
K
K
Chao
R L
RL
Pollock
E G
EG
D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure; D005306:Fertilization; D008854:Microscopy, Electron; D010063:Ovum / Q000648:ultrastructure*; D012617:Sea Urchins; D014817:Vitelline Membrane; D015053:Zygote / Q000648:ultrastructure*; D000516:alpha-Amylases / Q000378:metabolism
D000516:alpha-Amylases