Verhage H G, Mavrogianis P A, Boomsma R A, Schmidt A, Brenner R M, Slayden O V, Jaffe R C
Department of Obstetrics and Gynecology, University of Illinois at Chicago, 60612, USA.
Biol Reprod. 1997 Sep;57(3):525-31. doi: 10.1095/biolreprod57.3.525.
The objective of this study was to detect and characterize a secreted oviduct-specific glycoprotein (OGP) in the rhesus macaque (Macaca mulatta) and to compare the characteristics of this OGP to those previously characterized in baboons and women. Oviducts were obtained from untreated ovariectomized rhesus and from ovariectomized rhesus either treated with estradiol (E2) for 14 days or treated sequentially with E2 for 14 days and then with E2 plus progesterone (P4) for an additional 14 days. Segments of oviducts were either fixed for morphological analysis, cultured for OGP synthesis and release, or frozen for RNA analysis. The proteins present in the culture media were separated by one-dimensional SDS-PAGE, and OGP was detected on Western blots using polyclonal antibodies generated against the reduced form of baboon OGP or a 17-amino acid segment of the baboon core protein. Cross-reacting antigens were present in the 120-kDa region, identical to what was observed for baboon and human OGP. Indirect immunogold localization of OGP on thin sections demonstrated specific clustering of gold particles over the apical secretory granules of the secretory cells of the oviductal epithelium. A cDNA was generated using RT-PCR and 5' and 3' rapid amplification of cDNA ends (RACE), and sequenced. The total transcript was 2237 nucleotides in length plus a poly(A) tail. The largest open reading frame was 624 amino acids, which would produce a protein of 69.3 kDa. The nucleotide sequence was more than 95% identical to the nucleotide sequences of baboon and human OGP. Northern blots revealed a single message at 2.4 kilobases (kb) in oviduct samples obtained from E2-treated rhesus. This message was absent in oviducts obtained from untreated ovariectomized and from sequential E2 plus P4-treated rhesus macaques. In summary, the rhesus oviduct synthesizes and secretes an OGP in the presence of E2 that is immunologically and structurally similar to the baboon and human OGP. The presence of a highly homologous glycoprotein in several primates suggests a similar function for OGP in the reproductive process.
本研究的目的是检测和鉴定恒河猴（猕猴）分泌的输卵管特异性糖蛋白（OGP），并将该OGP的特性与先前在狒狒和人类中鉴定的特性进行比较。从未经处理的卵巢切除的恒河猴以及接受14天雌二醇（E2）处理或先接受14天E2处理、再接受14天E2加孕酮（P4）处理的卵巢切除的恒河猴中获取输卵管。输卵管片段要么固定用于形态学分析，要么培养用于OGP的合成和释放，要么冷冻用于RNA分析。培养基中存在的蛋白质通过一维SDS-PAGE分离，使用针对还原形式的狒狒OGP或狒狒核心蛋白的17个氨基酸片段产生的多克隆抗体在Western印迹上检测OGP。交叉反应抗原存在于120 kDa区域，与在狒狒和人类OGP中观察到的相同。OGP在薄片上的间接免疫金定位显示，金颗粒在输卵管上皮分泌细胞的顶端分泌颗粒上特异性聚集。使用RT-PCR和cDNA末端的5'和3'快速扩增（RACE）生成cDNA并进行测序。总转录本长度为2237个核苷酸加上一个聚腺苷酸尾巴。最大的开放阅读框为624个氨基酸，将产生一个69.3 kDa的蛋白质。核苷酸序列与狒狒和人类OGP的核苷酸序列有超过95%的同一性。Northern印迹显示，在从接受E2处理的恒河猴获得的输卵管样本中，在2.4千碱基（kb）处有一条单一的信息。在从未经处理的卵巢切除的恒河猴以及接受E2加P4顺序处理的恒河猴的输卵管中没有这条信息。总之，恒河猴输卵管在E2存在的情况下合成并分泌一种OGP，其在免疫学和结构上与狒狒和人类OGP相似。几种灵长类动物中存在高度同源的糖蛋白表明OGP在生殖过程中具有相似的功能。
