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Effects of sea urchin hyalin on gastrulation
Author(s) -
Razinia Ziba,
Carroll Edward J.,
CoyleThompson Cathy,
Oppenheimer Steven B.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1097-b
Subject(s) - strongylocentrotus purpuratus , sea urchin , gastrulation , human fertilization , embryo , microbiology and biotechnology , calcium , morphogenesis , chemistry , vitelline membrane , biology , hyaline , biophysics , anatomy , andrology , biochemistry , embryogenesis , botany , oocyte , medicine , organic chemistry , gene
The sea urchin hyaline layer is an extracellular matrix that develops soon after fertilization of Strongylocentrotus purpuratus eggs. The major constituent of this layer is the fibrillar glycoprotein hyalin that is essential for maintenance, organization and adhesion of cells during morphogenesis. Here, hyalin was isolated from newly fertilized S. purpuratus embryos via disruption of the vitelline envelope with dithiothreitol, followed by removal of the hyaline layer from the embryo surface with ice‐cold NaCl‐KCl. Hyalin was collected as the supernatant from the settled embryos and its purity was assessed on 2 percent agarose gels. 25 hr S. purpuratus embryos were incubated for 25 hrs at 15 C with normal sea water, hyalin/low calcium sea water, NaCl‐KCl/ low calcium sea water or low calcium sea water alone in 96 well microplates. The experiment was repeated 10 times. The hyalin‐treated and control samples were observed live and formaldehyde fixed using a Zeiss Axiolab photomicroscope. 87 percent (average) of all the controls developed complete archenterons. In the hyalin treated samples, 48 percent displayed no invagination, 49 percent showed incomplete, unattached archenterons, and 3 percent developed compete archenterons. Many studies, including this one, suggest that hyalin may be involved in gastrulation events such as initiation of invagination. The assay used here, because it allows precise quantification of the effects of substances on specific morphogenetic events, should help us understand exactly how molecules, such as hyalin, affect one specific event versus another (supported by NIH NIGMS SCORE, RISE, MARC, ITQ program and the Joseph Drown Foundation).