Kreis J , Bonß R , Feistel K , Vick P .

Overexpressed Cd63 mutant does not localize to late endosomes but blocks axis elongation: A Overexpressed RFP-tagged wt Cd63 (red) localizes to plasma membrane and endosomes in animal cap cells, whereas Cd63Y237A (green) accumulates only in the plasma membrane. B Control specimen depicted normal neurulation and normal sized notochords (n, outlined by black dashed line) in transversal section. Overexpression of cd63Y237A mRNA in dorsal lineage caused defects in neural tube closure. Sections revealed strongly expanded notochordal tissue (outlined by black dashed line) and widened floorplate tissue, as indicated by apparently unaltered apical constriction at the hinge points lateral of it (arrowheads). C Untreated specimen of st. 31 established wt AP axis extension and wt expression of pax6 indicating normal eye development. Unilateral overexpression (injected side marked by asterisk) of cd63Y237A mRNA resulted in embryos with shortened AP axis and reduced neural and optical expression of pax6. Quantification of axis phenotype. D Sagittal sections of st. 10 specimens revealed cd63 transcripts in deep dorsal mesoderm (arrowhead). With ongoing gastrulation, expression was detected in the posterior neuroectoderm and notochordal mesoderm (st. 11; arrowheads). During neurulation (st. 13, 16, 20) cd63 vanished from the neuroectoderm and became restricted to the notochord (arrowheads). Sagittal and transversal sections of late neurulas confirmed notochordal expression (arrowheads). a, anterior; AP, antero-posterior; co, control; d, dorsal; p, posterior; s, somites; st., stage; v, ventral; wt, wildtype.

Figure 1. Overexpressed Cd63 mutant does not localize to late endosomes but blocks axis elongation. A Overexpressed RFP-tagged wt Cd63 (red) localizes to plasma membrane and endosomes in animal cap cells, whereas Cd63Y237A (green) accumulates only in the plasma membrane.B Control specimen depicted normal neurulation and normal sized notochords (n, outlined by black dashed line) in transversal section. Overexpression of cd63Y237A mRNA in dorsal lineage caused defects in neural tube closure. Sections revealed strongly expanded notochordal tissue (outlined by black dashed line) and widened floorplate tissue, as indicated by apparently unaltered apical constriction at the hinge points lateral of it (arrowheads).C Untreated specimen of st. 31 established wt AP axis extension and wt expression of pax6 indicating normal eye development. Unilateral overexpression (injected side marked by asterisk) of cd63Y237A mRNA resulted in embryos with shortened AP axis and reduced neural and optical expression of pax6. Quantification of axis phenotype.D Sagittal sections of st. 10 specimens revealed cd63 transcripts in deep dorsal mesoderm (arrowhead). With ongoing gastrulation, expression was detected in the posterior neuroectoderm and notochordal mesoderm (st. 11; arrowheads). During neurulation (st. 13, 16, 20) cd63 vanished from the neuroectoderm and became restricted to the notochord (arrowheads). Sagittal and transversal sections of late neurulas confirmed notochordal expression (arrowheads).a, anterior; AP, antero-posterior; co, control; d, dorsal; p, posterior; s, somites; st., stage; v, ventral; wt, wildtype.

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