Immunoelectron microscopic localization of the neural recognition molecules L1, NCAM, and its isoform NCAM180, the NCAM-associated polysialic acid, beta1 integrin and the extracellular matrix molecule tenascin-R in synapses of the adult rat hippocampus.
We have investigated the possibility that morphologically different excitatory glutamatergic synapses of the "trisynaptic circuit" in the adult rodent hippocampus, which display different types of long-term potentiation (LTP), may express the immunoglobulin superfamily recognition molecules L1 and NCAM, the extracellular matrix molecule tenascin-R, and the extracellular matrix receptor constituent beta1 integrin in a differential manner. The neural cell adhesion molecules L1, NCAM (all three major isoforms), NCAM180 (the largest major isoform with the longest cytoplasmic domain), beta1 integrin, polysialic acid (PSA) associated with NCAM, and tenascin-R were localized by pre-embedding immunostaining procedures in the CA3/CA4 region (mossy fiber synapses) and in the dentate gyrus (spine synapses) of the adult rat hippocampus. Synaptic membranes of mossy fiber synapses where LTP is expressed presynaptically did not show detectable levels of immunoreactivity for any of the molecules/epitopes studied. L1, NCAM, and PSA, but not NCAM180 or beta1 integrin, were detectable on axonal membranes of fasciculating mossy fibers. In contrast to mossy fiber synapses, spine synapses in the outer third of the molecular layer of the dentate gyrus, which display postsynaptic expression mechanisms of LTP, were both immunopositive and immunonegative for NCAM, NCAM180, beta1 integrin, and PSA. Those spine synapses postsynaptically immunoreactive for NCAM or PSA also showed immunoreactivity on their presynaptic membranes. NCAM180 was not detectable presynaptically in spine synapses. L1 could not be found in spine synapses either pre- or postsynaptically. Also, the extracellular matrix molecule tenascin-R was not detectable in synaptic clefts of all synapses tested, but was amply present between fasciculating axons, axon-astrocyte contact areas, and astrocytic gap junctions. Differences in expression of the membrane-bound adhesion molecules at both types of synapses may reflect the different mechanisms for induction and/or maintenance of synaptic plasticity.
Schuster
T
T
Zentrum für Molekulare Neurobiologie, Universität Hamburg, Hamburg, Germany. Thomas.schuster@zmnh.uni-hamburg.de
Krug
M
M
Stalder
M
M
Hackel
N
N
Gerardy-Schahn
R
R
Schachner
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001253:Astrocytes / Q000648:ultrastructure; D001369:Axons / Q000648:ultrastructure; D018891:Dentate Gyrus / Q000737:chemistry / Q000648:ultrastructure; D016326:Extracellular Matrix Proteins / Q000032:analysis; D019084:Fluorescent Antibody Technique, Indirect; D017629:Gap Junctions / Q000737:chemistry / Q000648:ultrastructure; D006624:Hippocampus / Q000737:chemistry* / Q000166:cytology; D019012:Integrin beta1 / Q000032:analysis; D039841:Leukocyte L1 Antigen Complex; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D016253:Microscopy, Immunoelectron; D019599:Mossy Fibers, Hippocampal / Q000737:chemistry / Q000648:ultrastructure; D005810:Multigene Family; D009411:Nerve Endings / Q000737:chemistry / Q000648:ultrastructure; D009419:Nerve Tissue Proteins / Q000032:analysis*; D019006:Neural Cell Adhesion Molecules / Q000032:analysis / Q000737:chemistry; D009473:Neuronal Plasticity; D019580:Perforant Pathway / Q000737:chemistry / Q000648:ultrastructure; D020033:Protein Isoforms / Q000032:analysis*; D017966:Pyramidal Cells / Q000737:chemistry / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D012794:Sialic Acids / Q000032:analysis; D013048:Specimen Handling; D013569:Synapses / Q000737:chemistry* / Q000648:ultrastructure; D019063:Tenascin / Q000032:analysis
D000911:Antibodies, Monoclonal; D016326:Extracellular Matrix Proteins; D019012:Integrin beta1; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; D020033:Protein Isoforms; D012794:Sialic Acids; D019063:Tenascin; C021319:polysialic acid; C074638:tenascin R