Cloning and characterization of cDNA for syndecan core protein in sea urchin embryos.
The cDNA for the core protein of the heparan sulfate proteoglycan, syndecan, of embryos of the sea urchin Anthocidaris crassispina was cloned and characterized. Reverse transcription-polymerase chain reaction (RT-PCR) was used with total ribonucleic acid (RNA) from late gastrula stage embryos and degenerate primers for conserved regions of the core protein, to obtain a 0.1 kb PCR product. A late gastrula stage cDNA library was then screened using the PCR product as a probe. The clones obtained contained an open reading frame of 219 amino acid residues. The predicted product was 41.6% identical to mouse syndecan-1 in the region spanning the cytoplasmic and transmembrane domains. Northern analysis showed that the transcripts were present in unfertilized eggs and maximum expression was detected at the early gastrula stage. Syndecan mRNA was localized around the nuclei at the early cleavage stage, but was then found in the ectodermal cells of the gastrula embryos. Western blotting analysis using the antibody against the recombinant syndecan showed that the proteoglycan was present at a constant level from the unfertilized egg stage through to the pluteus larval stage. Immunostaining revealed that the protein was expressed on apical and basal surfaces of the epithelial wall in blastulae and gastrulae.
Tomita
K
K
Department of Regulation Biology, Faculty of Science, Saitama University, Urawa, Japan.
Yamasu
K
K
Suyemitsu
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D018076:DNA, Complementary; D007106:Immune Sera / Q000302:isolation & purification; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D011509:Proteoglycans / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012617:Sea Urchins / Q000196:embryology*; D017386:Sequence Homology, Amino Acid; D053668:Syndecan-1; D053667:Syndecans
D018076:DNA, Complementary; D007106:Immune Sera; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D012333:RNA, Messenger; C508242:Sdc1 protein, mouse; D053668:Syndecan-1; D053667:Syndecans