Distinct distribution of vimentin and cytokeratin in Xenopus oocytes and early embryos.
We report the identity of a major component of Triton-insoluble extracts from Xenopus oocytes and early embryos. In a previous paper we showed that an antibody, Z9, cross-reacts with two polypeptides from such extracts (Mr 56,000 and 57,000) as well as Xenopus vimentin. Direct microsequencing of the Mr 57,000 protein shows near identity of three tryptic fragments with regions of the predicted amino acid sequence of XCK1(8), a basic cytokeratin whose mRNA is known to be expressed in Xenopus oocytes. We have raised an antibody, CK7, against a fusion protein generated from this cDNA. The specificity of this antibody has been tested using 1- and 2-dimensional immunoblotting, which show that it is specific for the Mr 56,000 and 57,000 proteins, suggesting that these two proteins may be the products of two non-allelic XCK1(8) genes. The antibody does not cross-react with vimentin. We have used CK7 to follow the distribution of XCK1(8) throughout development by immunoblotting and immunocytochemistry. In larval stages, strong staining is seen in the notocord, the apical epithelia of the gut, the mesentery, and a few cells in the spinal cord. In oocytes and early embryos, two distinct intermediate filament (IF) networks can be distinguished: a cortical cytokeratin network, and a deeper vimentin one. In addition, the oocyte germ plasm stains with Z9 but not CK7. We propose that such distinct distributions of each IF protein reflect functional differences during early development.
Torpey
N P
NP
Wellcome Trust, University of Cambridge, UK.
Heasman
J
J
Wylie
C C
CC
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins / Q000737:chemistry; D007633:Keratins / Q000032:analysis*; D007814:Larva / Q000737:chemistry; D008969:Molecular Sequence Data; D009672:Notochord / Q000737:chemistry*; D009865:Oocytes / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D014746:Vimentin / Q000032:analysis*; D014981:Xenopus
D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014746:Vimentin; D007633:Keratins