Complex carbohydrates and complementary carbohydrate receptors on opposing cells may mediate neural cell interactions. The current application focuses on gangliosides, a major class of cell surface glycoconjugates in the central nervous system (CNS), as potential neural recognition molecules. High-affinity ganglioside conjugates were synthesized and used to identify a novel brain ganglioside receptor which may by involved in neural cell interactions. Purification and molecular characterization of this and related receptors will provide unprecedented molecular and immunological tools with which to probe the functions of gangliosides and related glycosphingolipids. Myelin-associated ganglioside receptor. A high-affinity, ganglioside- specific binding activity was recently identified on CNS myelin membranes. Its saccharide specificity and membrane distribution suggest that it may be involved in oligodendroglial-axonal recognition. The primary goal of the next grant period will be the isolation and molecular characterization of this binding activity. Various synthetic ganglioside conjugates have been prepared for use as high-affinity radioligands, affinity chromatography matrices, and photoaffinity probes. Molecular biological techniques will be used to determine the sequence of the myelin ganglioside receptor and to generate molecular and immunological tools with which to probe its distribution, developmental expression, and function. Synaptosomal ganglioside receptor. A second ganglioside-specific binding activity was found enriched on rat brain synaptosomes. Saccharide specificity and tissue/subcellular distribution studies of this second brain ganglioside receptor will be performed, leading to its purification and molecular characterization. Sulfoglucuronyl glycosphingolipids in peripheral nervous system (PNS) cell recognition. Schwann cells, the myelin forming cells of the PNS, were found to adhere specifically to purified sulfoglucuronyl glycolipids, anionic glycosphingolipids implicated in certain peripheral neuropathies. Receptors complementary to these lipids were detected on PNS membranes. They will be characterized for saccharide and tissue specificity, with the goal of purification, molecular characterization, and functional studies.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD014010-14
Application #
3312433
Study Section
Neurology C Study Section (NEUC)
Project Start
1980-04-01
Project End
1995-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
14
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Shaper, N L; Meurer, J A; Joziasse, D H et al. (1997) The chicken genome contains two functional nonallelic beta1,4-galactosyltransferase genes. Chromosomal assignment to syntenic regions tracks fate of the two gene lineages in the human genome. J Biol Chem 272:31389-99
Mahoney, J A; Schnaar, R L (1997) Multivalent ganglioside and sphingosine conjugates modulate myelin protein kinases. Biochim Biophys Acta 1328:30-40
Schnaar, R L; Longo, P; Yang, L J et al. (1996) Distinctive ganglioside patterns revealed by anti-ganglioside antibody binding to differentiating CG-4 oligodendrocytes. Glycobiology 6:257-63
Yang, L J; Zeller, C B; Schnaar, R L (1996) Detection and isolation of lectin-transfected COS cells based on cell adhesion to immobilized glycosphingolipids. Anal Biochem 236:161-7
Yang, L J; Zeller, C B; Shaper, N L et al. (1996) Gangliosides are neuronal ligands for myelin-associated glycoprotein. Proc Natl Acad Sci U S A 93:814-8
Schnaar, R L; Mahoney, J A; Swank-Hill, P et al. (1994) Receptors for gangliosides and related glycosphingolipids on central and peripheral nervous system cell membranes. Prog Brain Res 101:185-97
Mahoney, J A; Schnaar, R L (1994) Ganglioside-based neoglycoproteins. Methods Enzymol 242:17-27
White, T K; Schnaar, R L (1994) Solubilization of a membrane-associated protein from rat nervous system tissues which binds anionic glycolipids and phospholipids. Biochim Biophys Acta 1196:218-26
Needham, L K; Schnaar, R L (1993) Carbohydrate recognition in the peripheral nervous system: a calcium-dependent membrane binding site for HNK-1 reactive glycolipids potentially involved in Schwann cell adhesion. J Cell Biol 121:397-408
Needham, L K; Schnaar, R L (1993) The HNK-1 reactive sulfoglucuronyl glycolipids are ligands for L-selectin and P-selectin but not E-selectin. Proc Natl Acad Sci U S A 90:1359-63

Showing the most recent 10 out of 26 publications