N-type calcium channel in the pathogenesis of experimental autoimmune encephalomyelitis.
One of the family of voltage-gated calcium channels (VGCC), the N-type Ca(2+) channel, is located predominantly in neurons and is associated with a variety of neuronal responses, including neurodegeneration. A precise mechanism for how the N-type Ca(2+) channel plays a role in neurodegenerative disease, however, is unknown. In this study, we immunized N-type Ca(2+) channel α(1B)-deficient (α(1B)(-/-)) mice and their wild type (WT) littermates with myelin oligodendrocyte glycoprotein 35-55 and analyzed the progression of experimental autoimmune encephalomyelitis (EAE). The neurological symptoms of EAE in the α(1B)(-/-) mice were less severe than in the WT mice. In conjunction with these results, sections of the spinal cord (SC) from α(1B)(-/-) mice revealed a reduction in both leukocytic infiltration and demyelination compared with WT mice. No differences were observed in the delayed-type hypersensitivity response, spleen cell proliferation, or cytokine production from splenocytes between the two genotypes. On the other hand, Western blot array analysis and RT-PCR revealed that a typical increase in the expression of MCP-1 in the SC showed a good correlation with the infiltration of leukocytes into the SC. Likewise, immunohistochemical analysis showed that the predominant source of MCP-1 was activated microglia. The cytokine-induced production of MCP-1 in primary cultured microglia from WT mice was significantly higher than that from α(1B)(-/-) mice and was significantly inhibited by a selective N-type Ca(2+) channel antagonist, ω-conotoxin GVIA or a withdrawal of extracellular Ca(2+). These results suggest that the N-type Ca(2+) channel is involved in the pathogenesis of EAE at least in part by regulating MCP-1 production by microglia.
Tokuhara
Naoki
N
From the Department of Biochemistry and Molecular Pharmacology, Graduate School of Medicine, Chiba University, Chiba 260-8670; Department of Biology/Pharmacology, Discovery Research, Neuroscience Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Namiki
Kana
K
From the Department of Biochemistry and Molecular Pharmacology, Graduate School of Medicine, Chiba University, Chiba 260-8670.
Uesugi
Mai
M
Department of Genomics, Biomarkers and Personalized Medicine Core Function Unit, Tsukuba, Ibaraki 300-2635, Japan.
Miyamoto
Chihiro
C
Department of Tsukuba Discovery Research, KAN Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Ohgoh
Makoto
M
Department of Biology/Pharmacology, Discovery Research, Neuroscience Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Ido
Katsutoshi
K
Department of Biology/Pharmacology, Discovery Research, Neuroscience Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Yoshinaga
Takashi
T
Department of Global Drug Safety, Biopharmaceutical Assessment Core Function Unit, Eisai Company, Limited, Tsukuba, Ibaraki 300-2635, Japan.
Yamauchi
Toshihiko
T
Department of Biology/Pharmacology, Discovery Research, Neuroscience Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Kuromitsu
Junro
J
Department of Tsukuba Discovery Research, KAN Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Kimura
Sadao
S
From the Department of Biochemistry and Molecular Pharmacology, Graduate School of Medicine, Chiba University, Chiba 260-8670.
Miyamoto
Norimasa
N
Department of Tsukuba Discovery Research, KAN Product Creation Unit, Tsukuba, Ibaraki 300-2635, Japan.
Kasuya
Yoshitoshi
Y
From the Department of Biochemistry and Molecular Pharmacology, Graduate School of Medicine, Chiba University, Chiba 260-8670. Electronic address: kasuya@faculty.chiba-u.jp.
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020864:Calcium Channels, N-Type / Q000235:genetics / Q000378:metabolism*; D018932:Chemokine CCL2 / Q000096:biosynthesis* / Q000235:genetics; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D007962:Leukocytes / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008808:Mice, Inbred CBA; D008817:Mice, Mutant Strains; D017628:Microglia / Q000378:metabolism* / Q000473:pathology; D063308:Myelin-Oligodendrocyte Glycoprotein; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D013116:Spinal Cord / Q000378:metabolism* / Q000473:pathology; D020866:omega-Conotoxin GVIA / Q000494:pharmacology
D002121:Calcium Channel Blockers; D020864:Calcium Channels, N-Type; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D006023:Glycoproteins; D063308:Myelin-Oligodendrocyte Glycoprotein; D010446:Peptide Fragments; C445081:myelin oligodendrocyte glycoprotein (35-55); D020866:omega-Conotoxin GVIA; D002118:Calcium