Sprouting from chicken embryo dorsal root ganglia induced by nerve growth factor is specifically inhibited by affinity-purified antiganglioside antibodies.

Abstract

The involvement of gangliosides in processes related to nerve regeneration and sprouting has been demonstrated' recently. The type of interaction by which gangliosides may influence neuronal' sprouting was investigated in the present work.Affinity-purified rabbit anti-GMI antibodies were found to block the sprouting from dorsal root ganglia (DRG) of chicken embryo induced by nerve growth factor (NGF). Only a moderately inhibitory effect was produced by antibodies directed to GM2, suggesting a specific interaction between NGF and GM1. In spite of the low inhibitory effect ofanti-GM2 antibodies on sprouting, their binding to dissociated DRG cells was found to be higher than that of anti-GMl antibodies. These results emphasize the high resolution of the system and suggest that the interaction of NGF with GM1 is involved in the process of sprouting.Gangliosides are cell membrane-associated molecules that appear to have an important role in various cellular events including differentiation, growth, transformation, and regeneration (1-7). It has been demonstrated that administration of gangliosides intraperitoneally to denervated rats accelerates nerve regeneration (4-6). The addition ofgangliosides to dorsal root ganglia (DRG) and neuroblastoma cells cultured in vitro has been found to promote neurite outgrowth from these cells (8). These studies suggested that neuronal gangliosides play a role as mediator molecules in trophic interactions. Indeed, we recently have found that specific antibodies to gangliosides block neuritic outgrowth from goldfish regenerating retinal explants (7).Nerve growth factor (NGF) from mouse salivary gland is known to induce sprouting (9-11). We have now extended our investigation of the role of gangliosides by studying sprouting from DRG induced by NGF. We have used liposomes containing gangliosides as an agent for affinity purification of antiganglioside antibodies. We found that addition of affinity-purified anti-GM1 antibodies to cultured DRG blocks the sprouting induced by J3-NGF. By contrast, antibodies against GM2 had only a limited effect, in spite of their higher extent of binding to dissociated DRG cells. It is suggested from the present study that gangliosides, in particular GM1, are involved in the process of sprouting from DRG induced by NGF. MATERIALS AND METHODS Gangliosides. These were. extracted from whole bovine brains with chloroform/methanol and' then subjected to alkali treatment and phase partition. The gangliosides were further purified by column chromatography on DEAE-Sephadex A-25 with elution by chloroform/methanol/0.8 M sodium acetate, 30: 60: 8 (vol/vol) (12). The ganglioside preparations were found to be completely devoid of contaminating proteins detectable by amino acid analysis.Preparation of Antibodies. Outbred rabbits were obtained from the breeding colony ofthe Animal Breeding Center ofthe Weizmann Institute of Science. The rabbits were immunized with highly purified mixed gangliosides (5 mg) complexed noncovalently to methylated bovine serum a...

Passive Immunization with Anti-Ganglioside Antibodies Directly Inhibits Axon Regeneration in an Animal Model

“…Longitudinal studies will be required to address the question whether clearance of Abs from circulation allows regeneration of inhibited nerve fibers in this model. Overall, this concept of inhibition of axon regeneration through gangliosides is also supported by previous work showing that Abs against GM1 ganglioside can inhibit regeneration in nonmammalian neurons (Schwartz and Spirman, 1982;Sparrow et al, 1984).…”

Development and survival of neurons in dissociated fetal mesencephalic serum-free cell cultures: II. Modulatory effects of gangliosides

“…In fact, although neurite outgrowth has not been evaluated in this study, GM 1 has been reported to facilitate neurite outgrowth in cells both with and without an absolute neuronotrophic requirement for survival and/or neurite extension (Byrne et al, 1983;Ferrari et al, 1983;Facci et al, 1984;KatohSemba et al, 1984;Leon et al, 1984;Matta et al, 1986). Interestingly, in chicken DRG cells in vitro, gangliosides potentiate NGF-induced neurofilament expression and maintenance (Doherty et al, 1985) while antibodies to GM1 block in a dose-dependent manner the NGF-induced neurite outgrowth (Schwartz and Spirman, 1982). In SB21B2 cells, there occurs during ganglioside-induced differentiation a significant increase in the expression of mRNA for tubulin (Rybak et al, 1983).…”

Morphological and phenotypic changes of nociceptive primary afferent neurons following neurotmesis and perineural capsaicin treatment

“…Hence, a peripheral nerve lesion induces fundamental changes in the chemical phenotype of chemosensitive primary sensory neurons, which express the capsaicin/TRPV1 receptor: these neurons which, in the intact animal do not bind the GM1 ganglioside change their phenotype and, similarly to large DRG neurons, turn into GM1 binding ganglion cells. GM1 plays a critical role in the mechanisms of the trophic actions of growth factors, in particular nerve growth factor (NGF, (Schwartz and Spirman, 1982, Leon et al, 1984, Mutoh et al, 1998)which, in turn, plays a pivotal role in the regulation of the capsaicin sensitivity of DRG neurons . Since in rat sensory ganglion neurons the availability of NGF, and consequently the expression of the capsaicin/TRPV1 receptor is critically dependent on the retrograde axonal transport , Aguayo and White, 1992, Bevan and Winter, 1995, it seemed worthwhile to initiate further studies to reveal possible changes in the expression of the TRPV1 receptor following different types of peripheral nerve lesions.…”