1997 Fiscal Year Final Research Report Summary
Molecular cloning of IL-3 receptor-associated antigen
| Project/Area Number |
08457198
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Neurology
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| Research Institution | National Institute of Neuroscience, National Center of Neurology and Psychiatry |
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Principal Investigator |
TABIRA Takeshi National Institute of Neuroscience, Department of Demyelinating Disease and Aging.Head, 神経研究所・神経研究所疾病研究第6部, 部長 (80112332)
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| Co-Investigator(Kenkyū-buntansha) |
KONISHI Yoshihiro kawasaki Medical School, Department of Neuropathology, Assistant Professor, 神経病理学, 講師 (90170290)
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| Project Period (FY) |
1996 – 1997
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| Keywords | interleukin-3(IL-3) / IL-3 receptor / anti-IL-3 receptor / IL-3 receptor-associated antigen / cholinergic neurons / neurotrophic factor / molecular clonig |
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| Research Abstract |
We previously reported that interleukin-3(IL-3) acts as a neurotrophic factor for murine septum-derived cholinergic neurons. And it is interesting to know how IL-3 acts on these neurons via the specific receptors. F9 monoclonal antibody recognizes IL-3 receptor-associated antigen(IL-3RAA) which is different from alpha of beta subunit of IL-3 receptor. Surprisingly, IL-3RAA is distributed in most of large neurons throughout the central nervous system(CNS), although alpha and beta subunit immunoreacrtivities are relatively restricted to cholinergic neurons of the basal forebrain. To understand the mechanisms involved in the effect of IL-3 in the CNS,cloning IL-3RAA is very useful.
F9 antibody recognized three plasma membrane proteins at 150,100 and 50 kDa on Western blot analysis. The 50 kDa of F9-reactive protein was most abundant. It was isolated, purified, then its two partial amino acid sequences were identified. Another antibody raised against the partial amino acid also recognized similar neurons. According to these amino acid sequences, two pairs of oligonucleotide primers were synthesized, used for RT-PCR amplification of mRNAs isolated from SN6 cells which are cholinergic and positive for F9 antibody. Amplified DNA fragments were inserted into T vector of pUC18, cloned, then their DNA sequences were determined.
We obtained 25 clones of the DNA frabments. Three were unknown and hybridized with 1.6 and 3.0 kb mRNA.The 3.0 kb mRNA was expressed specifically in the CNS.We are now analyzing the DNA sequences of the total length of these clones. Progress in clarifying a molecule recognized by F9 antibody will provide useful infromation for understanding the mechanisms involved in cholinergic neurotrophic function of IL-3 in the CNS.
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