Immunochemical Analysis of Arylsulfatase Accumulation in Sea Urchin Embryos: extracellular matrix/arylsulfatase/sea urchin embryo/tissue‐specific gene products/sea urchin embryo/in situ hybridization

Qing Yang, Paul D. Kingsley, David J. Kozlowski, Robert C. Angerer, Lynne M. Angerer

Research output: Contribution to journalArticle

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Abstract

We have determined the expression pattern of arylsulfatase in embryos of the sea urchin Strongylocentrotus purpuratus. Polyclonal antibodies raised against a fusion protein containing sequences encoded by SpARSI (Yang et al., 1989, Dev. Biol. 135: 53–61, 1989) detect several peptides of 65–70 kD on immunoblots. Treatment with glycopeptidase F shows that at least one of these peptides is modified by N‐linked glycosylation, which accounts for some of the peptide diversity. We have also identified a second arylsulfatase gene (SpARSII) whose sequence is highly similar to ARS, a gene expressed in the Hemicentrotus pulcherrimus embryo. Arylsulfatase activity is detectable in unfertilized eggs, in which only SpARSII mRNA can be detected. Both SpARSI and SpARSII mRNAs increase greatly in abundance during embryogenesis accompanied by parallel changes in arylsulfatase activity and immunoreactivity. Immunohistochemistry with the anti‐SpARSI antibody shows that arylsulfatase accumulates primarily along the apical surface of the aboral ectoderm of pluteus larvae, and to a lesser extent along portions of oral ectoderm. At earlier stages, the protein is more uniformly distributed along all presumptive ectoderm, reflecting a more uniform mRNA distribution. Treatment of embryos with glycine‐EDTA, which dissociates but does not lyse cells of the embryo, releases virtually all enzymatic activity and all immunoreactive protein. Embryos cultured in sulfate‐free sea water, which arrest at gastrula stage, show normal accumulation and secretion of peptide detected with the SpARSI antibody.

Original languageEnglish (US)
Pages (from-to)139-151
Number of pages13
JournalDevelopment, Growth & Differentiation
Volume35
Issue number2
DOIs
StatePublished - Apr 1993
Externally publishedYes

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ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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