title: IgG anti-GalNAc-GD1a antibody inhibits the voltage-dependent calcium channel currents in PC12 pheochromocytoma cells.
abstract: We investigated the effects of IgG anti-GalNAc-GD1a antibodies, produced by immunizing rabbits with GalNAc-GD1a, on the voltage-dependent calcium channel (VDCCs) currents in nerve growth factor (NGF)-differentiated PC12 pheochromocytoma cells. VDCCs currents in NGF-differentiated PC12 cells were recorded using the whole-cell patch-clamp technique. Immunized rabbit serum that had a high titer of anti-GalNAc-GD1a antibodies inhibited the VDCCs currents in the NGF-differentiated PC12 cells (36.0+/-9.6% reduction). The inhibitory effect of this serum was reversed to some degree within 3-4 min by washing with bath solution. Similarly, application of purified IgG from rabbit serum immunized with GalNAc-GD1a significantly inhibited the VDCCs currents in PC12 cells (30.6+/-2.5% reduction), and this inhibition was recovered by washing with bath solution. Furthermore, the inhibitory effect was also observed in the GalNAc-GD1a affinity column binding fraction (reduction of 31.1+/-9.85%), while the GalNAc-GD1a affinity column pass-through fraction attenuated the inhibitory effect on VDCCs currents. Normal rabbit serum and normal rabbit IgG did not affect the VDCCs currents in the PC12 cells. In an immunocytochemical study using fluorescence staining, the PC12 cells were stained using GalNAc-GD1a binding fraction. These results indicate that anti-GalNAc-GD1a antibodies inhibit the VDCCs currents in NGF-differentiated PC12 cells.
authors.affiliation: Department of Pharmacotherapics, Showa Pharmaceutical University, Machida, Tokyo 194-8543, Japan.
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Nakatani
authors.forename: Takumi
authors.initials: T
authors.lastname: Nagaoka
authors.forename: Sayako
authors.initials: S
authors.lastname: Hotta
authors.forename: Iku
authors.initials: I
authors.lastname: Utsunomiya
authors.forename: Hiide
authors.initials: H
authors.lastname: Yoshino
authors.forename: Tadashi
authors.initials: T
authors.lastname: Miyatake
authors.forename: Keiko
authors.initials: K
authors.lastname: Hoshi
authors.forename: Kyoji
authors.initials: K
authors.lastname: Taguchi
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation; D004553:Electric Conductivity; D005732:Gangliosides / Q000276:immunology*; D007074:Immunoglobulin G / Q000494:pharmacology*; D007158:Immunologic Techniques; D020932:Nerve Growth Factor / Q000494:pharmacology; D016716:PC12 Cells / Q000378:metabolism* / Q000473:pathology; D011817:Rabbits; D051381:Rats; D013194:Staining and Labeling
chemical_list: D015220:Calcium Channels; D005732:Gangliosides; D007074:Immunoglobulin G; C055253:IV(4)-galactosyl-N-acetylganglioside GD1a; D020932:Nerve Growth Factor