ISOLATION OF SECONDARY MESENCHYME CELL SURFACE SPECIFIC ANTIGEN FROM ECHINODERM EMBRYOS, AND ANALYSIS OF ITS ROLE AND GENE EXPRESSION IN EMBRYOGENESIS
| Project/Area Number |
09680719
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Developmental biology
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
KATOW Hideki GRADUATE SCHOOL OF SCIENCE, TOHOKU UNIVERSITY, PROFESSOR, 大学院・理学研究科, 教授 (30111610)
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| Co-Investigator(Kenkyū-buntansha) |
KURAISHI Ritsu GRADUATE SCHOOL OF SCIENCE, TOHOKU UNIVERSITY, ASSISTANT, 大学院・理学研究科, 助手 (60195526)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | ECHINODERM EMBRYO / SECONDARY MESENCHYME CELL / CELL SURFACE-SPECIFIC ANTIGEN / MONOCLONAL ANTIBODY / DELAMINATION |
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| Research Abstract |
Delamination of migratory embryonic cells during embryogenesis is a rather general morphogenetic process in multicellular animals. Thus it is basic and important process needed to be elucidated its molecular basis. In sea urchin embryos the primary mesenchyme (PMC) and the secondary mesenchyme cells (SMC) delaminate from the vegetal plate ectoderm and at the invaginating tip of endoderm, respectively. This process has been suggested to be associated with alteration of cell/cell adhesion property. In this regard, the present research project was aimed to find out if there is any cell surface-specific protein that shows the change of its way of presence on the delaminating cell during delamination process. We have successfully produced a monoclonal antibody (mAb) that recognizes a cell surface-specific protein. The protein was expressed spatio-temporally during mesenchyme cell ingression. Its relative molecular mass was 35kDa and monomeric (P35). P35 was detected on the basal surface mesenchyme cells during ingression that is the period the basal lamina components are degraded to make access for mesenchyme cells to the blastocoel. According to immunochemistry P35 had no epitope that was recognized by antibodies raised against the known matrix metalloproteases. Isolation of the protein from embryo needed detergent-treatment suggesting P35 is a membrane integrated or strongly membrane-bound component. Microinjection of mAb did not cause morphogenetic disturbances. We interpret this consequence that mAb may not bind to active site of P35 which is rather common observation for monoclonal antibodies. Thus to further understand the biological function, we will raise polyclonal antibodies. To analyze gene expression, the polyclonal antibodies will be used as a probe to screening cDNA library constructed in λgt11 phage.
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Report
(4 results)
Research Products
(3 results)
