Inhibition of memory consolidation by antibodies against cell adhesion molecules after active avoidance conditioning in zebrafish.
Cell adhesion molecules are expected to play an important role in long-term storage of information in the central nervous system. Several of these glycoproteins, such as NCAM, L1, and the ependymins, express the HNK-1 carbohydrate structure, which is known to be involved in cell-cell and cell-matrix interactions. To investigate the contribution of the HNK-1 epitope and the secretory glycoproteins ependymins to memory formation in zebrafish (Brachydanio rerio), we developed an active avoidance conditioning paradigm. Zebrafish were trained in a shuttle-box to cross a hurdle, to avoid mild electric shocks following a conditioning light signal. One hour after acquisition of the task, zebrafish were injected intracerebroventricularly with monoclonal antibodies against the HNK-1 epitope or polyclonal antibodies against ependymins. Control fish received immunoglobulins G (IgGs) from nonimmune rat serum or the monoclonal antibody C183 against an unrelated cell-surface protein of the cyprinid brain. Two days later, injected zebrafish were tested for recall, and for quantitative evaluation a retention score (RS), ranging from 1.0 for immediate recall to 0.0, indicating no saving, was calculated. The average RS of anti-HNK-1-injected fish (RS = 0.30) and anti-ependymin-injected fish (0.24) were significantly different from the RS of uninjected fish (0.77), of controls injected with nonimmune serum IgGs (0.68), of C183-injected controls (0.78), and of overtrained fish injected with anti-HNK-1 antibodies (0.81). Anti-HNK-1 and anti-ependymin antibodies were characterized by Western blot analyses of subcellular brain fractions and immunohistochemical staining of the zebrafish optic tectum. Our data suggest that the antibodies influence cell recognition events at synaptic membranes and/or associated intracellular signaling cascades, and thereby block memory consolidation.
Pradel
G
G
Biotechnology Center, Justus-Liebig-University, Giessen, Germany.
Schachner
M
M
Schmidt
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000954:Antigens, Surface / Q000276:immunology; D001362:Avoidance Learning / Q000502:physiology*; D015153:Blotting, Western; D001923:Brain Chemistry / Q000502:physiology; D003213:Conditioning, Psychological / Q000502:physiology; D000939:Epitopes / Q000276:immunology; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins / Q000276:immunology; D008568:Memory / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000276:immunology; D019006:Neural Cell Adhesion Molecules / Q000276:immunology*; D009473:Neuronal Plasticity / Q000502:physiology*; D013347:Subcellular Fractions / Q000737:chemistry; D015238:Sulfotransferases / Q000276:immunology; D015027:Zebrafish / Q000502:physiology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000939:Epitopes; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; C031632:ependymins; C109318:HNK-1 sulfotransferase; D015238:Sulfotransferases