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The morphological events during the self-reassembly of the sea urchin embryo in culture

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Title The morphological events during the self-reassembly of the sea urchin embryo in culture
Names Hamada, Spencer Hiroshi (creator)
Harris, Patricia (advisor)
Date Issued 1975-08-15 (iso8601)
Note Graduation date: 1976
Abstract The gastrula of the sea urchin Strongylocentrotus purpuratus
was dissociated into single cells. In culture these cells appeared
apolar, but reassembled into quasi-normal embryos composed of
structures with polarized cells. In this study emphasis was placed on
those morphological events coincident with the reestablishment of a
polar orientation within the epithelial cells. Using both time-lapse
dark-field-phase microscopy and electron microscopy, the major
morphological events followed in detail include: (a) the migration
of lysosome-like vesicles to specific margins within the epithelial
cells; (b) the migration of the epithelial cells during the re-construction
of the epithelium: and (c) the reformation of the cell junctions between
the epithelial cells.
Evidence indicates that:
(1) The orientation resulting from the migration of the
lysosome-like vesicles is transient and may not determine any lasting
polarity of the epithelial cell. The sequential migration of the
lysosome-like vesicles first to the epithelial cell to cell interface,
then to the basal margin and finally to the apical margin of the
epithelial cell was coincident with and apparently in response to the
presence of cell debris often trapped at these sites. The Lysosome-like
vesicles formed tight clusters along the epithelial cellular margins
where phagocytosis of cell debris and resorption of specialized
structures occurred. In contrast, the mesenchyme cell had smaller
numbers of phagosomes and lysosome-like vesicles and was observed
to participate to a more limited degree in phagocytosis and resorption.
(2) The orientation resulting from the formation of a leading edge
during the migration of the epithelial cell is transient. The sequence
of cytological events associated with epithelial cell migration included
the formation of: (a) hyaloplasmic blebs along the cell free
margin; (b) filopodia and microvilli; and (c) lamellae with a number
of lamellipodia at the leading edge. In contrast, the mesenchyme cell
formed filopodia on a smaller scale and was observed to participate
in migration only to a limited degree. Coincident with the cessation
of epithelial cell migration was the enlargement of the lamella between
the epithelial cells into a hyaloplasmic sheet which forms the blastocoel
wall.
Embryonic cells from reassembling aggregates were cultured as
monolayers on glass treated with an adhesion enhancing factor derived from the incubation medium. Cords of cells, composed of both
epithelial and mesenchyme cells migrate away from the aggregate
and leave in their wake clusters of cells. Epithelial cells were found
at the leading margins of these cell cords and were observed forming
cytoplasmic blebs, filopodia and hyaloplasmic leading edges during
the migration of the cell cord. No apparent coordinate orientation
was detectable within the epithelial cells of the migrating cell cord
or stationary cell cluster.
A hypothesis is presented suggesting a role for resorption
during epithelial cell migration.
(3) Cell contacts formed during reaggregation become more
adhesive as development proceeds. The sequence of fine structural
events associated with cell contact formation include the formation
of (a) a non--junctional cell apposition, which was characterized by
parallel apposed membranes; (b) an incipient continuous junction,
which was characterized by parallel apposed membranes; and (c) a
continuous junction, which was characterized by the presence of an
electron dense region both superjacent and subjacent to the plasma
membrane at the site of the junction.
The cell appositions between premigratory cells were easier
to disrupt mechanically than the later forming continuous junctions.
The forming continuous junction was first focal in extent and later
zonal in girth around the apical margin of the epithelial cell.
Concurrent with the formation of the strongly adhesive
continuous junction by the lamella was a cessation of epithelial cell
migration, followed by an expression of an apical-basal polarity
within the epithelial cell.
A hypothesis is presented suggesting a role for cell contacts
during the coordinated orientation of the epithelia during the formation
of the blastocoel wall.
Genre Thesis/Dissertation
Topic Sea urchin embryo
Identifier http://hdl.handle.net/1957/45587

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