Supporting Online Material


Individual Cell Migration Serves as the Driving Force for Optic Vesicle Evagination
Martina Rembold, Felix Loosli, Richard J. Adams, Joachim Wittbrodt

Supporting Online Material

This supplement contains:
Materials and Methods
Fig. S1
References
Movies S1 to S14

This file is in Adobe Acrobat PDF format.

Movie S1
Gross morphology of optic vesicle evagination in wild-type. RPCs: green (rx3::GFP), neural plate cells: red, (histoneH2B-mRFP). A single confocal plane from a time lapse recording shows the convergence of the eye field as the neural keel forms and the subsequent evagination of the optic vesicles. The eye field remains wide as compared to the neural keel before evagination.

Movie S2
Failure of evagination in eyeless. Mutant RPCs (green, rx3::GFP) continuously move towards the midline. They fail to evaginate and a neural keel is formed instead.

Movie S3
Frontal view of a 4D reconstruction of cells tracked during optic vesicle evagination in wildtype; dorsal is to the top. RPCs (green) of the ventro-medial eye field remain stationary while the lateral eye field and the lateral forebrain cells (magenta) converge towards the midline.

Movie S4
Dorsal view of a 4D reconstruction of optic vesicle evagination in wild-type showing stronger convergence of lateral forebrain cells (magenta) as compared to RPCs (green), a process that keeps the eye field wide.

Movie S5
Dorsal view of a 4D reconstruction of morphogenesis in eyeless. RPCs (green) converge at the same rate towards the midline as forebrain cells (magenta), resulting in the formation of a neural keel instead of eyes.

Movie S6
Cell morphology during optic vesicle morphogenesis in wild-type and eyeless visualized by rx3::mYFP. A single confocal plane shows the dynamic shape changes RPCs undergo during evagination in wild-type. In eyeless, the mutant RPCs elongate and incorporate into the forming epithelium of the neural tube.

Movie S7
Mosaic analysis in the wild-type background. Wild-type cells (red, membrane-mRFP) in a wildtype eye field (green, rx3::mYFP) participate normally in optic vesicle evagination.

Movie S8
Mosaic analysis in the eyeless background. Rescue of optic vesicle evagination by wild-type cells (red), mutant cells (green) form part of the neural keel.

Movie S9
4D reconstruction of rescued optic vesicle evagination. Wild-type cells (red, asterisk) located in the medial eye field move through the mutant tissue (green) and participate in the forming optic vesicle. Only one half of the eye field is shown.

Movie S10
Cell migration. A cluster of approx. 4 wild-type cells (yellow dot) migrates through the surrounding rx3-mutant tissue and becomes part of the forming optic vesicles.

Movie S11
Single cell migration. A single wild-type cell (red, asterisk) migrates through the mutant tissue in el (green) and eventually becomes part of the forming optic vesicle.

Movie S12
Cell morphology of RPCs during migration visualized by rx3::mYFP. A wild-type RPC (green, rx3::mYFP) in an unlabelled wild-type eye field elongates and extends a lamellipodium as it migrates to the right towards the evaginating optic vesicles. The movie shows a maximum-intensity projection of several confocal planes.

Movie S13
Neighbour exchange during migration. Cell 1 is initially not a direct neighbour of cell 2 but migrates over it during evagination.

Movie S14
RPC morphology in the eye field. A cell extends filopodia and lamellipodia as it moves towards the midline.

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