SEARCH FOR NOVEL ANGIOGENIC FACTORS IN THE OVARY

研究了控制血管平滑肌细胞增殖的新因素。通过多种层析从牛卵泡液中分离出促进血管平滑肌细胞增殖的多肽。测定了这些多肽的部分氨基酸序列，并根据氨基酸序列用简并寡核苷酸探针分离了它们的cDNA克隆。在卵泡液中发现了几个活性组分。从肝素琼脂糖层析的1.0M氯化钠组分中鉴定出两种生长因子：中期因子和多营养素是VSMC的生长因子。活性组份进一步用肝素琼脂糖柱层析纯化为均一组分，经SDS PAGE鉴定为80 kDa的单一多肽。测定了该多肽的部分氨基酸序列，分离到编码该多肽的cDNA克隆，并测定了其核苷酸序列。对DNA数据库的搜索显示，该多肽是F-spaindin，被认为参与了胎儿的分化。

New factors controlling vascular smooth muscle cell proliferation have been investigated.Polypeptides promoting smooth muscle cell proliferation were isolated from bovine follicular fluid by a several kinds of chromatographies. Partial amino acid sequences of these polypeptides were determined and their cDNA clones were isolated by degenerative oligonucleotide probes based on the amino acid sequences. A several active fractions were found in the follicular fluid. From 1.0M NaCl fraction from Heparin Sepharose chromatography, two growth factors, midkine and pleiotrophin were identified as growth factors of VSMC.Another activity was eluted from DEAE Sepharose chromatography at 0.3M NaCl concentration. The active fraction was further purified to homogeneity by Heparin Sepharose chromatography and was characterized on SDS PAGE as a 80kDa single polypeptide. Partial amino acid sequence of the polypeptide was determined and a cDNA clone encoding the polypeptide was isolated and the nucleotide sequence was determined. Search on the DNA data bank revealed the polypeptide was F-spondin, which is believed to participate in the differenciation of fetus.