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Antisera to an axolemma-enriched fraction have anti-axon and anti-myelin effects in vitro
20 2 second, high intensity sound (Ii0 db at 2900 hz, VI-I min.) 2 hours daily for 4 days. The femurs and tibias were dissected from the animals, bone marrow cells separated and suspended in media. In vitro migration assays were employed using blind well chambers. Thymus supernatant from 3 day old mice was used as the attractant in the bottom well separated from the bone marrow cells in the top well by Nucleopore membranes having 5 ~m diameter pores. After 90 minutes incubation, cells migrating to the bottom chamber were collected, counted and compared to controls. Preliminary studies suggest a measurable increase in the number of migrating bone marrow cells in the stressed females suggesting that acute stress may enhance pre-thymic stem cell production in the bone marrow. Future studies will evaluate the effects of acute versus chronic stress on pre-thymic cell populations. (Supported by NIH grant AG 04384)
ANTISERA FO AN AXOLEMMA-ENRICHED FffACTION HAVE ANTI-AXON AND ANTI~YELIN EFFECTS IN VITRO Dennis N. Bourdette L, Fredrick J. Sell ~ , John W. Bigbee 2, George H. DeVries ~, Charles K. Meshul ~ , and Harish C. Agraw~l ~ IVA Medical Center and Department of Neurology, Oregon Health Sciences University, Portland, OR; 2 " " " V 3Department of Biochemistry, Medical College of Virglnla, Richmond, A; Departments of Pediatrics and Neurology, Washington University School of Medicine, St. Louis Children's Hospital, St. Louis, MO. Rabbit antisera to an axolenwa-enriched fraction (AEF) of rat brain contain antibodies that bind to several axolemmal proteins and immunohlstochemically stain the surface of peripheral nerve axons (Bigbee et al, J Neurolmmunol 5 (1983):191). We tested 5 antlsera to AEF for antiaxon and anti-myelin--effects by applying them to mouse spinal cord-dorsal root ganglia (SC-DRG) explant cultures. All 5 antisera inhibited neurite outgrowth from the SC and DRG explants when applied to the c Jltures from the time of explantatlon. The antlsera destroyed outgrowth axons when applied to mature SC-DRG cultures. Electron microscopic examination of anterior horn ceils and DRG neurons exposed to the antisera revealed an increase in free ribosomes and neurofilaments, consistent with an axotomy reaction. Three of the antisera to AEF inhibited myelination when applied to the cultures from the time of explantation but t ~ s e antisera did not demyelinate mature c,Jltures. Two of the antlsera had no anti-myelin effects. An antiserum to the chloroform-methanol insoluble proteins of myelin caused both myellnatlon inhibition and demyelination. The 3 antisera to AEF that caused myelination inhibition contained antibodies to galactocerebroslde while the 2 antisera that were negative for myelination inhibition did not, suggesting that the anti-myelin effect of these antisera was mediated by antibodies to galactocerebroside. Antisera to AEF can destroy axons and inhibit neurite outgrowth and some antisera may inhibit myelination in vitro. These in vitro studies may be relevant to some human diseases in which irmnune reac---tio---n-s'toaxons may occur. Supported by the Veterans Administration and by NIH grants NS13464, NS19414, NSIO821, and NS15408.
ANTISERA FO AN AXOLEMMA-ENRICHED FffACTION HAVE ANTI-AXON AND ANTI~YELIN EFFECTS IN VITRO Dennis N. Bourdette L, Fredrick J. Sell ~ , John W. Bigbee 2, George H. DeVries ~, Charles K. Meshul ~ , and Harish C. Agraw~l ~ IVA Medical Center and Department of Neurology, Oregon Health Sciences University, Portland, OR; 2 " " " V 3Department of Biochemistry, Medical College of Virglnla, Richmond, A; Departments of Pediatrics and Neurology, Washington University School of Medicine, St. Louis Children's Hospital, St. Louis, MO. Rabbit antisera to an axolenwa-enriched fraction (AEF) of rat brain contain antibodies that bind to several axolemmal proteins and immunohlstochemically stain the surface of peripheral nerve axons (Bigbee et al, J Neurolmmunol 5 (1983):191). We tested 5 antlsera to AEF for antiaxon and anti-myelin--effects by applying them to mouse spinal cord-dorsal root ganglia (SC-DRG) explant cultures. All 5 antisera inhibited neurite outgrowth from the SC and DRG explants when applied to the c Jltures from the time of explantatlon. The antlsera destroyed outgrowth axons when applied to mature SC-DRG cultures. Electron microscopic examination of anterior horn ceils and DRG neurons exposed to the antisera revealed an increase in free ribosomes and neurofilaments, consistent with an axotomy reaction. Three of the antisera to AEF inhibited myelination when applied to the cultures from the time of explantation but t ~ s e antisera did not demyelinate mature c,Jltures. Two of the antlsera had no anti-myelin effects. An antiserum to the chloroform-methanol insoluble proteins of myelin caused both myellnatlon inhibition and demyelination. The 3 antisera to AEF that caused myelination inhibition contained antibodies to galactocerebroslde while the 2 antisera that were negative for myelination inhibition did not, suggesting that the anti-myelin effect of these antisera was mediated by antibodies to galactocerebroside. Antisera to AEF can destroy axons and inhibit neurite outgrowth and some antisera may inhibit myelination in vitro. These in vitro studies may be relevant to some human diseases in which irmnune reac---tio---n-s'toaxons may occur. Supported by the Veterans Administration and by NIH grants NS13464, NS19414, NSIO821, and NS15408.