| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2006: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
Relaxin (RLX) was discovered as one of the pregnancy hormones, but it is active at not only genital organ but also brain and heart and its versatile roles are widely noticed. Therefore, an antibody to detect RLX is needed. RLX, which resembles insulin in structure, comes from proRLX, a precursor of RLX, after removal of the C-peptide, and is composed of A and B chains connected by an S-S bond. The A chain has also an S-S bond. Due to the presence of these S-S bonds, it was difficult to synthesize the antibody which could recognize RLX by immunity with a fragment of RLX. On the other hand, significantly, a half-life of the proRLX is longer than that of RLX in vivo. Thus, we made an anti-C-peptide human RLX antibody to detect local existence of RLX and established the immunity-organisms staining method using this for as a primary antibody. The anti-C-peptide antibody was prepared by immunity of a rabbit using a C-peptide chain of human proRLX which was produced in an E. coli. We found th
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at RLX have growth promotion action of a follicle of an early stage in a culture experiment of a Human follicle cortex graft (J Clin Endocrinol Metab 90:516, 2005). We applied this antibody to the examination why RLX in a human follicle in an early stage was localized. Dyeing was detected in ovum and granule membrane cell of primordial, primary, and second follicles. On the other hand, the localization in a follicle of LGR7, an RLX receptor, was recognized by in situ hybridizaton and immunity dyeing in flat granule membrane cells in the human primordial follicle, granule membrane cells in a second follicle. From these results, we can concluded that our first clarification of the activity of RLX toward early-stage development of human follicle via both autocrine and paracrine mechanisms with its receptor. This suggested that RLX is possibly one of the maturity initiating factors for the human primordial follicle. The anti-C-peptide RLX antibody provided in this study was useful for examination of localization of RLX production in human and a pig. We also perform the total synthesis of human RLX (H2) to examine follicle maturity action and mechanism of RLX more and more. Less
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