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A novel microdissection model for probing cellular interactions. I. alpha methyl mannose blocks the cellular interaction
Author(s) -
CoyleThompson Catherine,
Askari Tina,
Mkrtchian Mary,
Portillo Georgina,
Manukyan Mariam,
Abramian Sarineh,
Qubrosi Mirey,
Makhoul Elias,
Frederick Robert,
Allen Jenieke,
Dreyfuss Justin,
Oppenheimer Steven B.
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.695.3
Subject(s) - blastocoel , pronase , mannose , gastrulation , microbiology and biotechnology , chemistry , biology , embryo , biochemistry , embryogenesis , trypsin , enzyme
Adhesion between the archenteron tip and blastocoel roof in the NIH designated sea urchin embryo model is a cellular interaction that has interested investigators for over a century, but its molecular basis is not understood. Here we microdissect the two components of this cellular interaction and use alpha methyl mannose to map molecular groups that may be involved in mediating the cellular interaction. Whole 48–54 hour fixed Lytechinus pictus embryos were washed three times and then dissected in pH 8.0 artificial sea water (ASW), 15°C. The roofs of the blastocoel and the tips of the archenteron were put together and observed to stick together. In over 50 separate trials using bound sets of roof and archenterons attached together, either 0.2 M or 1.0 M alpha methyl mannose was added to the pieces. The pieces remained together. However in 100% of the cases, manually separating the pieces in alpha methyl mannose at either molarity resulted in blocking the rebinding of the blastocoel roofs and archenteron tips. Re‐binding occurred in the absence of the sugar. The results suggest that alpha methyl mannose binding receptors are involved in mediating the cellular interaction. These studies offer a novel approach to map glycans and glycan binding partners that may be functionally important. By microdissecting the components of cellular interactions out of the embryo proper, these components can be probed in pristine media away from factors in intact embryos that could confuse results (Supported by NIH NIGMS SCORE S0648680, MARC, RISE, the Joseph Drown Foundation, the Sidney Stern Memorial Trust, and CSU Northridge Biology Full Immersion Research Experience (FIRE) course funding).