| Project/Area Number |
09680715
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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 | HOKKAIDO UNIVERSITY |
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Principal Investigator |
MITA Koichi Grad.School of Sci., Hokkaido Univ., Inst., 大学院・理学研究科, 助手 (50281845)
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| Co-Investigator(Kenkyū-buntansha) |
片桐 千明 北海道大学, 大学院・理学研究科, 教授 (90000827)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Vitelline coat / Hatching gland cell / Hatching enzyme / Oviduct / Oviductin / エゾアカガエル |
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| Research Abstract |
1. To analyze the mechanisms of the differentiation of the hatching gland cells (HGCs) in Xenopus laevis, we have recombined various portions of the dorsal mesoderm of the early gastrula with ectoderm. Explants and control embryos were fixed after reaching stage 25. Hatching gland cells are identified with an antibody against hatching enzyme (alpha-UVS.2 antibody). Results are summarized as follows : (1) Surface ectoderm of presumptive head region can be induced to form HGCs by the underlying mesoderm. (2) Anterior region of the mesoderm is a candidate for the inducer of HGCs. (3) Inducing activity is potential until at least late gastrula stage. (4) Posterior region of an ectoderm can be differentiate into HGCs at mid gastrula stage, but lost its ability in the later stage. (5) At late gastrula stage, only anterior portion of the presumptive head region has an ability to differentiate into HGCs.
2. To isolate Bufo oviductin cDNA, we produced two degenerate PCR primers, which correspond to N-terminal amino acid sequences of Bufo oviductin (for a forward primer) and serine-specific amino acid sequences (for a reverse primer). PCR template was produced from the total RNA of pars recta oviduct of Bufo japonicus. PCR products were cloned into a plasmid vector pGEM-Easy Vector and sequenced. This clone had an insert of 570 bp, and the predicted amino acid sequence exhibited significant homology (57%) with Xenopus oviductin, although the entire coding region was not covered. Now we are trying to isolate clones including the entire coding region of Bufo oviductin from a cDNA library.
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