The polysialic acid moiety of the neural cell adhesion molecule is involved in intraretinal guidance of retinal ganglion cell axons.
We have characterized the antigen recognized by mab10, a monoclonal antibody that has been shown to modify outgrowth of thalamic and cortical axons in vitro, and investigated the influence of this antibody on axonal growth in the chicken retina in vivo. Immunopurification, peptide sequencing, and biochemical characterization proved the epitope recognized by mab10 to be polysialic acid (PSA), associated with the neural cell adhesion molecule (NCAM). Intravitreal injections of antibody-secreting hybridoma cells were combined with whole-mount studies using the fluorescent tracer 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (DiI). Pathfinding at the optic fissure was affected, resulting in a failure of axons to exit into the nerve. Misprojections also occurred in more peripheral areas of the retina; however, axons eventually oriented toward the center. Similar projection errors were observed after enzymatic removal of PSA by injecting endoneuraminidase N (endo N). Quantitative measurements of the optic nerve diameter as well as the width of the optic fiber layer confirmed that many axons failed to leave the retina and grew back in the optic fiber layer of the retina. Our findings suggest that NCAM-linked PSA is involved in guiding ganglion cell axons in the retina and at the optic fissure.
Monnier
P P
PP
Department of Ophthalmology, University of Tübingen, Schleichstrasse 12, Tübingen, D-72076, Germany.
Beck
S G
SG
Bolz
J
J
Henke-Fahle
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001369:Axons / Q000502:physiology*; D001923:Brain Chemistry; D002450:Cell Communication; D002642:Chick Embryo; D000939:Epitopes; D006026:Glycoside Hydrolases / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism*; D009900:Optic Nerve / Q000196:embryology* / Q000473:pathology; D051381:Rats; D012160:Retina / Q000196:embryology*; D012165:Retinal Ganglion Cells / Q000502:physiology*; D012794:Sialic Acids / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D019006:Neural Cell Adhesion Molecules; D012794:Sialic Acids; C021319:polysialic acid; D006026:Glycoside Hydrolases; C090534:endo-alpha-sialidase