Identification of vimentin and novel vimentin-related proteins in Xenopus oocytes and early embryos.
We have made antibodies against fusion proteins of Xenopus vimentin. We show for the first time the distribution of vimentin in larval stages, where it is found in cells of mesenchymal origin, and in radial glial cells. In sections of Xenopus oocytes and early embryos, immunocytochemistry reveals the presence of an extensive cytoplasmic network, distributed in an animal-vegetal gradient. Germ plasm stains particularly strongly. The form of the IF proteins in this network is unusual. In immunoblot experiments the anti-vimentin antibodies detect a number of distinct proteins. We have identified those that are the products of the two known vimentin genes, by injection of synthetic mRNA transcribed from cloned vimentin cDNAs into oocytes, followed by two-dimensional Western blotting. This has demonstrated unambiguously that one Xenopus vimentin, Vim1, is present in oocytes and early embryos. However, two other immunoreactive proteins detected in Triton extracts of oocytes and early embryos are not the products of Vim1, since depletion of vimentin mRNA by antisense oligonucleotide injection has no effect on the synthesis of these proteins. These results suggest that novel IF-like proteins are expressed in Xenopus oocytes and early embryos.
Torpey
N P
NP
Department of Zoology, University of Cambridge, UK.
Heasman
J
J
Wylie
C C
CC
D000818:Animals; D000911:Antibodies, Monoclonal; D015152:Blotting, Northern; D003593:Cytoplasm / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004625:Embryo, Nonmammalian / Q000737:chemistry*; D005260:Female; D015151:Immunoblotting; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000276:immunology; D008648:Mesoderm / Q000737:chemistry; D016253:Microscopy, Immunoelectron; D009865:Oocytes / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014746:Vimentin / Q000032:analysis*; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins; D011993:Recombinant Fusion Proteins; D014746:Vimentin