Monoclonal antibodies as probes for the differentiation of the mouse trophectoderm
Monoclonal antibodies as probes for the differentiation of the mouse trophectoderm
The aim of this study was to use monoclonal antibodies to identify and characterise antigens specific to the trophectoderm of the preimplantation mouse blastocyst and in this way to further describe the molecular features of the trophectoderm phenotye and mechanisms regulating trophectoderm biogenesis. In attempts to identify antigens unique to the trophectoderm, an intrasplenic immunisation technique was developed and using blastocysts as the immunogen, hybridomas secreting trophectoderm specific antibodies were identified.
Trophectoderm specific antigen characterisation was carried out for an antigen recognised by a monoclonal antibody coded 283D3. This antibody was raised in the mouse against the rabbit yolk sac and has previously been shown to recognise a high molecular weight antigen (330-380kDa) in the rabbit, mouse and rat yolk sac, endoderm and proximal tubule epithelia. In the trophectoderm, 283D3 was found to recognise a 350kDa antigen localised to the apical trophectoderm surface and apical intracellular sites. Ultrastructurally, it was shown that at least some epitopes are displayed on the outer face of the plasma membrane, and that intracellularly the antigen is localised to variously sized apical non-coated endocytic vesicles and some cytosolic sites. Antigen expression was also detected in late morulae of the same age post hCG as early blastocysts (not earlier stages), but only at the apical surface. By manipulating cavitation indirectly using the anti-uvomorulin antibody ECCD-1, and directly using the drug ouabain, intracellular antigen epxression was shown to be dependent on cavitation. While onset of expression at both sites occurred only from the 6th cell cycle onwards it was independent of cytokinesis and also occurred independently of cell flattening and cell-cell adhesion at compaction and beyond.
While the function of the 283D3 antigen was not determined, presence of the antibody seemed to retard development and a role as a nutrient or growth factor receptor has been speculated. Through the use of the 283D3 monoclonal antibody probe, a novel feature of the trophectoderm phenotype has been identified. Trophectoderm biogenesis seems to include the late appearance of an apical surface and vesicularly localised antigen and it has been revealed that the assembly of some epithelial components is regulated by cavitation while some occur independent of this event.
University of Southampton
1993
Butler, Elizabeth Ann
(1993)
Monoclonal antibodies as probes for the differentiation of the mouse trophectoderm.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
The aim of this study was to use monoclonal antibodies to identify and characterise antigens specific to the trophectoderm of the preimplantation mouse blastocyst and in this way to further describe the molecular features of the trophectoderm phenotye and mechanisms regulating trophectoderm biogenesis. In attempts to identify antigens unique to the trophectoderm, an intrasplenic immunisation technique was developed and using blastocysts as the immunogen, hybridomas secreting trophectoderm specific antibodies were identified.
Trophectoderm specific antigen characterisation was carried out for an antigen recognised by a monoclonal antibody coded 283D3. This antibody was raised in the mouse against the rabbit yolk sac and has previously been shown to recognise a high molecular weight antigen (330-380kDa) in the rabbit, mouse and rat yolk sac, endoderm and proximal tubule epithelia. In the trophectoderm, 283D3 was found to recognise a 350kDa antigen localised to the apical trophectoderm surface and apical intracellular sites. Ultrastructurally, it was shown that at least some epitopes are displayed on the outer face of the plasma membrane, and that intracellularly the antigen is localised to variously sized apical non-coated endocytic vesicles and some cytosolic sites. Antigen expression was also detected in late morulae of the same age post hCG as early blastocysts (not earlier stages), but only at the apical surface. By manipulating cavitation indirectly using the anti-uvomorulin antibody ECCD-1, and directly using the drug ouabain, intracellular antigen epxression was shown to be dependent on cavitation. While onset of expression at both sites occurred only from the 6th cell cycle onwards it was independent of cytokinesis and also occurred independently of cell flattening and cell-cell adhesion at compaction and beyond.
While the function of the 283D3 antigen was not determined, presence of the antibody seemed to retard development and a role as a nutrient or growth factor receptor has been speculated. Through the use of the 283D3 monoclonal antibody probe, a novel feature of the trophectoderm phenotype has been identified. Trophectoderm biogenesis seems to include the late appearance of an apical surface and vesicularly localised antigen and it has been revealed that the assembly of some epithelial components is regulated by cavitation while some occur independent of this event.
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Published date: 1993
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Local EPrints ID: 462420
URI: http://eprints.soton.ac.uk/id/eprint/462420
PURE UUID: e4d51267-2da7-4f5f-99b5-16accb4a8345
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Date deposited: 04 Jul 2022 19:07
Last modified: 04 Jul 2022 19:07
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Author:
Elizabeth Ann Butler
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