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J Histochem Cytochem. 2014 Feb;62(2):145-56. doi: 10.1369/0022155413511619. Epub 2013 Oct 23.

KSGal6ST is essential for the 6-sulfation of galactose within keratan sulfate in early postnatal brain.

Author information

1
Section of Pathophysiology and Neurobiology, Research, National Center for Geriatrics and Gerontology, (HH, SO, KU), Obu, Japan.

Abstract

Keratan sulfate (KS) comprises repeating disaccharides of galactose (Gal) and N-acetylglucosamine (GlcNAc). Residues of Gal and GlcNAc in KS are potentially modified with sulfate at their C-6 positions. The 5D4 monoclonal antibody recognizes KS structures containing Gal and GlcNAc, both 6-sulfated, and has been used most extensively to evaluate KS expression in mammalian brains. We previously showed that GlcNAc6ST1 is an enzyme responsible for the synthesis of the 5D4 epitope in developing brain and in the adult brain, where it is induced after injury. It has been unclear which sulfotransferase is responsible for Gal-6-sulfation within the 5D4 KS epitope in developing brains. We produced mice deficient in KSGal6ST, a Gal-6-sulfotransferase. Western blotting and immunoprecipitation revealed that all 5D4-immunoreactivity to proteins, including phosphacan, were abolished in KSGal6ST-deficient postnatal brains. Likewise, the 5D4 epitope, expressed primarily in the cortical marginal zone and subplate and dorsal thalamus, was eliminated in KSGal6ST-deficient mice. Disaccharide analysis showed the loss of Gal-6-sulfate in KS of the KSGal6ST-deficient brains. Transfection studies revealed that GlcNAc6ST1 and KSGal6ST cooperated in the expression of the 5D4 KS epitope in HeLa cells. These results indicate that KSGal6ST is essential for C-6 sulfation of Gal within KS in early postnatal brains.

KEYWORDS:

5D4; developing brain; extracellular matrix; glycosaminoglycan; keratan sulfate; phosphacan; sulfotransferase

PMID:
24152993
PMCID:
PMC3902094
DOI:
10.1369/0022155413511619
[Indexed for MEDLINE]
Free PMC Article

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