Weaver granule neurons are rescued by calcium channel antagonists and antibodies against a neurite outgrowth domain of the B2 chain of laminin.
The weaver mutation impairs migration of the cerebellar granular neurons and induces neuronal death during the first two weeks of postnatal life. To elucidate the molecular mechanisms for the impaired neuronal migration, we investigated the rescue mechanisms of the weaver (wv/wv) granule neurons in vitro. We found that Fab2 fragments of antibodies against a neurite outgrowth domain of the B2 chain of laminin enhanced neurite outgrowth and neuronal migration of the weaver granule neurons on a laminin substratum and in the established cable culture system. The rescue of the weaver granule neurons by antibodies against the B2 chain of laminin may result from the neutralizing effect of these antibodies against the elevated B2 chain levels of the weaver brain. The L-type calcium channel blocker, verapamil (1-5 microM), also rescued the weaver granule neurons. High concentrations of MK-801 (10-20 microM), a glutamate receptor antagonist and voltage-gated calcium channel blocker, rescued the weaver granule neurons similar to verapamil, but low concentrations of MK-801 (1 microM) had no rescue effect. Simultaneous patch-clamp studies indicated that the weaver granule neurons did not express functional N-methyl-D-aspartate receptors further indicating that the rescue of the weaver granule neurons by MK-801 resulted from its known inhibition of voltage-gated calcium channels. The present results indicate that antibodies against the B2 chain of laminin, verapamil, and high concentrations of MK-801 protect the weaver granule neurons from the otherwise destructive action of the weaver gene. Thus, both the laminin system and calcium channel function contribute to the migration deficiency of the weaver granule neurons.
Liesi
P
P
Laboratory of Molecular and Cellular Neurobiology, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, Maryland 20852, USA. liesi@helix.nih.gov
Wright
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002465:Cell Movement; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D007797:Laminin / Q000037:antagonists & inhibitors / Q000502:physiology*; D039841:Leukocyte L1 Antigen Complex; D051379:Mice; D008817:Mice, Mutant Strains; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010455:Peptides / Q000494:pharmacology; D011817:Rabbits; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors / Q000378:metabolism; D014633:Valine / Q000031:analogs & derivatives / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology; D020866:omega-Conotoxin GVIA
D000911:Antibodies, Monoclonal; D002121:Calcium Channel Blockers; D007797:Laminin; D039841:Leukocyte L1 Antigen Complex; D019006:Neural Cell Adhesion Molecules; D010455:Peptides; D016194:Receptors, N-Methyl-D-Aspartate; D016291:Dizocilpine Maleate; C056639:2-amino-5-phosphopentanoic acid; D020866:omega-Conotoxin GVIA; D014700:Verapamil; D014633:Valine