2003 Fiscal Year Final Research Report Summary
Studies on laminin domains involved in the epithelial organogenesis.
| Project/Area Number |
13670023
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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 |
General anatomy (including Histology/Embryology)
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| Research Institution | Kitasato University |
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Principal Investigator |
KADOYA Yuichi Kitasato Univ., School of Medicine, Assistant Professor, 医学部, 講師 (10185887)
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| Co-Investigator(Kenkyū-buntansha) |
NOMIZU Motoyoshi Hokkaido Univ., Graduate School of Environmental Earth Science, Associate Professor, 大学院・地球環境科学研究科, 助教授 (00311522)
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| Project Period (FY) |
2001 – 2003
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| Keywords | antibody / cell adhesion / laminin / organogenesis / peptide |
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| Research Abstract |
Laminins are multifunctional extracellular glycoproteins found in the basement membranes (BMs) of various organs. At least 15 laminin isoforms have been identified. Among them, laminin-1 and -10 were found to co-localize in the epithelial BMs of embryonic mouse submandibular gland (SMG), suggesting their crucial rotes for epithelial organogenesis. In order to identify functional domains of laminin-1 and -10, we screened more than 50 cell-adhesive 12-mer synthetic laminin-1 and -10 peptides known to support HT1080 cell adhesion, in the organ cultures of rudimental SMG or hair follicles, or a cell adhesion assay using epithelial liver cell line. As a result, we identified four functional laminin peptides, AG73 peptide (RKRLQVQLSIRT) and A5G77f peptide (LVLFLNHGH), which inhibited epithelial branching morphogenesis : and A5G33 peptide (ASKAIQVFLLAG) and A5G65 peptide (HQNMGSVNVSVG), which supported epithelial cell spreading.
Based on these findings, attempts were made to establish antibodies against the functional laminin peptides. AG73, A5G77f, and A5G33 peptides were synthesized and conjugated to keyhole limpet hemocyanin (KLH). Rabbits were injected with the peptide-KLH 3 to 5 times at 2-week intervals. In some experiments, guinea pigs were used instead of rabbits. None of these peptides, however, raised sufficient amount of specific antibodies. We therefore synthesized a multimeric form of A5G77f and immunized rabbits with it. An antiserum R3 showed affinity for A5G77f 100-1000 times as high as that for A5G77fT, a control scramble peptide for A5G77f. Using an organ culture and cell adhesion assays, we are now testing biological functions of R3.
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[Publications] Hayashi, K., Mochizuki, M., Nomizu, M., Uchinuma, E., Yamashina, S., Kadoya, Y.: "Inhibition of Hair Follicle Growth by a Laminin-1 G-Domain Peptide, RKRLQVQLSIRT, in an Organ Culture of Isolated Vibrissa Rudiment."J.Invest.Dermatol.. 118. 48-52 (2002)
Description
「研究成果報告書概要(和文)」より
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[Publications] Hayashi, K., Mochizuki, M., Nomizu, M., Uchinuma, E., Yamashina, S., Kadoya, Y.: "Inhibition of Hair Follicle Growth by a Laminin-1 G-Domain Peptide, RKRLQVQLSIRT, in an Organ Culture of Isolated Vibrissa Rudiment."J.Invest.Dermatol.. 118. 48-52 (2002)
Description
「研究成果報告書概要(欧文)」より
