1992 Fiscal Year Final Research Report Summary
Development of Neuronal Labelling and applied techniques by using Bacterial Toxins
| Project/Area Number |
03557002
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
神経解剖学
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| Research Institution | UNIVERSITY OF TSUKUBA |
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Principal Investigator |
OKADO Noboru UNIV. TSUKUBA Dep. Anat Associate Professor, 基礎医学系, 助教授 (50060140)
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| Co-Investigator(Kenkyū-buntansha) |
HOSOYA Yasuhiro UNIV. TSUKUBA Dep. Anat Associate Professor, 基礎医学系, 助教授 (60100145)
KUDO Norio UNIV. TSUKUBA Dep. Physiol Professor, 基礎医学系, 教授 (60014239)
HAYASHI Hideo UNIV. TSUKUBA Dep. Microbiol Professor, 基礎医学系, 教授 (40033203)
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| Project Period (FY) |
1991 – 1992
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| Keywords | Neuronal labelling / Bacterial toxin / Cholera toxin / E. Coli / 脊髄 / 運動ニューロン / コレラ毒素 / 大腸菌毒素 |
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| Research Abstract |
In order to develop neuronal labelling methods we tried to use bacterial toxins. Toxins were injected into the posterior iliotibial muscle of newly hatched chicken. Following a few days survival period chickens were sacrificed, and sections of the spinal cord were immunohistochemically examined by the use of an antibody against each toxin. Motoneurons and primary afferent fibers were clearly labelled with Cholera toxin B subunit and E. coil enterotoxin B subunit, which have strong affinity to one of the neuronal surface antigens, GMI ganglioside. Those toxins were not differentiated with monoclonal antibodies. By the use of shigella toxin or tetanus toxin neuronal structures were not labelled. For a double labelling method with bacterial toxins it was necessary to find out other bacterial toxins than those of cholera or E. coli. F frament of leukocidin, part of Staphylococcus aureus toxin appeared to the best candidate, because it has a strong affinity to GMI ganglioside. To produce a large amount of leukocidin we tried to introduce leikocidine gene into E. coli. However, E. coli could not produce leikocidine. into E. coli. However, E. coli could not produce leikocidine.
Although bacteral toxins are highly sensitive for neuronal labelling, there is a serious drawback. That was diffusions of bacterial toxins from the injectio sites. To eliminate this drawback cholera toxin conjugated with latex beads can be sucessfully applied for neuronal labelling.
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