Expression of glypican-4 in haematopoietic-progenitor and bone-marrow-stromal cells

Biochem J. 1999 Dec 15;344 Pt 3(Pt 3):937-43.

Abstract

Heparan sulphate proteoglycans and the extracellular matrix of bone-marrow-stromal cells are important components of the microenvironment of haematopoietic tissues and are involved in the interaction of haematopoietic stem and stromal cells. Previous studies have emphasized the role of heparan sulphate proteoglycan synthesis by bone-marrow-stromal cells. In the present study we describe the expression of glypican-4 (GPC-4), belonging to the glypican family, in bone-marrow-stromal cells and haematopoietic-progenitor cells of human and murine origin. Expression of GPC-4 was shown on the mRNA-level by reverse transcription-PCR and Northern blot analysis. Amplification products were cloned and sequenced, to confirm these results. To analyze the expression of GPC-4 on the protein level, polyclonal antibodies against selected peptides were raised in rabbits. Western blot analysis showed expression of GPC-4 as a heparan sulphate proteoglycan in the human haematopoietic-progenitor cell line TF-1 and normal human bone marrow. These results were confirmed by FACS analysis of TF-1 cells. Furthermore, GPC-4-positive progenitor cells and stromal cells were enriched from normal human bone marrow by magnetic-cell sorting and analysed by confocal laser-scanning microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Bone Marrow Cells / metabolism*
  • Cloning, Molecular
  • Flow Cytometry
  • Glypicans
  • Hematopoiesis
  • Hematopoietic Stem Cells / metabolism*
  • Heparan Sulfate Proteoglycans / genetics*
  • Heparan Sulfate Proteoglycans / metabolism
  • Humans
  • Immunohistochemistry
  • Mice
  • Microscopy, Confocal
  • Molecular Sequence Data
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stromal Cells / metabolism

Substances

  • GPC4 protein, human
  • Glypicans
  • Gpc4 protein, mouse
  • Heparan Sulfate Proteoglycans
  • RNA, Messenger