2001 Fiscal Year Final Research Report Summary
Electorophysiological and biochemistrical study on protecture mechaninism of odontoblasts identified with specific monoclonal antibodies
| Project/Area Number |
10470404
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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 |
Conservative dentistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
IKEDA Hideharu Tokyo Medical and Dental University, Dental Hospital, Lecturer, 歯学部・附属病院, 講師 (20222896)
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| Co-Investigator(Kenkyū-buntansha) |
SUDA Hideaki Tokyo Medical and Dental University, Graduate School, Pulp Biology and Endodontics, Professor, 大学院・医歯学総合研究科, 教授 (00114760)
SUNAKAWA Mitsuhiro Tokyo Medical and Dental University, Dental Hospital, Associate Professor, 歯学部・附属病院, 助教授 (30179288)
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| Project Period (FY) |
1998 – 2001
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| Keywords | odontoblast / monoclonal antibody / patch-clamp / cohole-cell / hystatin / gap-junction / electrical synapse |
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| Research Abstract |
We showed the functional link between odontoblasts (OB) and other type of pulpal cells (PC) beneath the odontoblastic layer.
1. Isolation and in vitro immunization : From cat and human dental pulp, outer pulpal cells including odontoblasts were freshly isolated. After fusion and screening, monoclonal antibodies that labeled odontoblasts were chosen.
2. Patch-clamp technique : Cells having a monopolar process and an oval cell body were used for patch-clamp recording. We succeeded to record single-channel activity and then nystatin-perforated whole-cell configuration was performed. Simply linear relation between current and voltage was obtained.
3. Dye-coupling : When lucifer yellow was injected into one of two types of cells with iontophoresis, the other type of cell was also labeled. But uncoupled cells, even though they were close to the cells, were not labeled, suggesting that lucifer yellow did not leak to the outer-cellular space and labeled the cell membrane, but spread into different kind of cells through dye-coupling.
4. Gap-junction : EM study showed that the couplings mentioned above were gap-junctions. After we confirmed that lucifer yellow injected in an OB or a PC transferred to the other type of cell(s), cells were fixed and used for EM study. Between two types of cells, gap-junctions were always observed.
5. Electrophysiological evidence of electrical coupling : Current could pass bidirectionally through the structure confirmed above morphologically. However, asymmetric transduction was observed.
These results demonstrate that OBs and PCs are functionally linked and work as a network and that OBs work as a sensor of external stimuli.
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