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Use of post‐fertilization aggregates of sea urchin embryos from denuded eggs to study self‐adhesive properties of the hyaline layer: glycosidases
Author(s) -
Jarvis Krystal,
Idoni Brian,
Dreyfuss Justin,
CoyleThompson Catherine,
Carroll Edward,
Oppenheimer Steven,
HutchinsCarroll Virginia
Publication year - 2010
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.24.1_supplement.695.6
Subject(s) - hyaline , human fertilization , embryo , sea urchin , vitelline membrane , botany , biology , andrology , amylase , chemistry , anatomy , microbiology and biotechnology , enzyme , oocyte , biochemistry , medicine
Vitelline envelope free sea urchin eggs do not form fertilization envelopes; however, the coat free embryos form aggregates (or clumps) as their exposed hyaline layers develop (Epel, Exp Cell Res 61(1):69–70, 1970). Here we describe a study using that property to probe hyaline layer interactions. De‐jellied Lytechinus pictus eggs were treated with 0.02 M dithiothreitol, pH 9.1, to remove vitelline envelopes, fertilized, washed 3X with artificial sea water (ASW), incubated at 5°C for 45 min and then incubated for 75 min at 17°C with no additions or with 40–9800 units/ml ASW of independently characterized α‐amylase, β‐amylase or β‐glucosidase. In 12 replicate experiments using eggs from 48 female sea urchins, counting at least 31,400 single embryos and clumps for all experiments, only β‐glucosidase caused disaggregation of clumps (p < 0.0001 compared with controls). Beta amylase caused single embryos to adhere to each other forming clumps (p < 0.03 compared with controls) and α‐amylase had no significant effect on the embryos (p > 0.19 compared with controls). The results suggest that the self‐aggregating property of coat free, fertilized embryos is a simple and effective system to probe hyaline layer‐hyaline layer interactions and that β‐glucosidic linked polysaccharides may be involved in these interactions (supported by NIH NIGMS SCORE S0648680, MARC, RISE, the Joseph Drown Foundation, the Sidney Stern Memorial Trust and CSU Northridge FIRE course funding).