Accessing the embryo interior without microinjection.
For decades it has been assumed that in order to insert macromolecules into the embryo blastocoel for numerous experimental purposes, microinjection was required. Microinjection, however, can be only performed on a few embryos at a time, thus precluding many studies that could involve large populations of embryos. Laser scanning confocal microscopy, with its optical sectioning advantage, showed that fluorochrome-labeled macromolecular lectins and bovine albumin enter the blastocoel of living, swimming sea urchin embryos following a period of incubation without microinjection. A procedure is also described that shows macromolecular entry is substantially accelerated in low calcium seawater. The information gained from this study should greatly facilitate experiments on entire populations of millions of embryos at a time that require access of macromolecules to the embryo interior.
Latham
V H
VH
Center for Cancer and Developmental Biology, California State University, Northridge 91330-8303, USA.
Martinez
A L
AL
Cazares
L
L
Hamburger
H
H
Tully
M J
MJ
Oppenheimer
S B
SB
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D007202:Indicators and Reagents; D037102:Lectins / Q000378:metabolism; D046911:Macromolecular Substances; D008845:Microinjections; D018613:Microscopy, Confocal; D012617:Sea Urchins / Q000196:embryology* / Q000378:metabolism* / Q000648:ultrastructure; D012623:Seawater; D012710:Serum Albumin, Bovine / Q000378:metabolism
D007202:Indicators and Reagents; D037102:Lectins; D046911:Macromolecular Substances; C039217:fluorescein isothiocyanate bovine serum albumin; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate; D002118:Calcium