Haworth J D, Rollyson M K, Silva P, McIntush E W, Niswender G D
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523, USA.
Biol Reprod. 1998 Jan;58(1):169-74. doi: 10.1095/biolreprod58.1.169.
To investigate expression of monocyte chemoattractant protein-1 (MCP-1) in the ovine corpus luteum, a partial cDNA was produced by reverse transcription-polymerase chain reaction. This cDNA was 89% identical to that reported for bovine MCP-1 mRNA. In experiment 1, steady-state concentrations of mRNA encoding MCP-1 were measured in pools of luteal tissue collected on Days 3, 6, 9, 12, and 15 of the estrous cycle (estrus = O; n = 4/day). There were no differences in mRNA concentrations for MCP-1 among any of the days studied (p = 0.43). In experiment 2, midluteal-phase corpora lutea were collected from ewes at 0 (untreated), 2, 4, 8, and 16 h after administration of a luteolytic dose of prostaglandin F2alpha (PGF2alpha; n = 4/time point). Concentrations of MCP-1 mRNA were undetectable in untreated controls, were detectable at 2 h post-treatment, had increased 4 and 8 h after administration of PGF2alpha when compared to those at 2 h (p < 0.05), and were decreased 16 h after administration of PGF2alpha when compared to those at 4 h (p < 0.05). In situ hybridization for MCP-1 mRNA combined with immunocytochemical labeling of tissue inhibitor of metalloproteinase-1 (TIMP-1) in large luteal cells was used to determine whether the steroidogenic cells that have PGF2alpha receptors express MCP-1 mRNA in response to PGF2alpha. Messenger RNA encoding MCP-1 and TIMP-1 were not colocalized, indicating that MCP-1 was not expressed by large steroidogenic luteal cells during luteolysis.
为研究单核细胞趋化蛋白-1(MCP-1)在绵羊黄体中的表达情况,通过逆转录-聚合酶链反应制备了部分cDNA。该cDNA与已报道的牛MCP-1 mRNA的cDNA有89%的同源性。在实验1中,测定了发情周期第3、6、9、12和15天(发情期=0;每组n = 4只/天)收集的黄体组织样本中编码MCP-1的mRNA的稳态浓度。在所研究的任何一天中,MCP-1的mRNA浓度均无差异(p = 0.43)。在实验2中,在给母羊注射溶黄体剂量的前列腺素F2α(PGF2α)后0(未处理)、2、4、8和16小时收集黄体中期的黄体(每组n = 4只/时间点)。未处理的对照组中未检测到MCP-1 mRNA的浓度,处理后2小时可检测到,与2小时时相比,PGF2α给药后4小时和8小时MCP-1 mRNA浓度升高(p < 0.05),与4小时时相比,PGF2α给药后16小时MCP-1 mRNA浓度降低(p < 0.05)。采用MCP-1 mRNA原位杂交结合大黄体细胞中金属蛋白酶组织抑制剂-1(TIMP-1)的免疫细胞化学标记,以确定具有PGF2α受体的类固醇生成细胞是否对PGF2α产生反应而表达MCP-1 mRNA。编码MCP-1和TIMP-1的信使RNA没有共定位,表明在黄体溶解过程中,大的类固醇生成黄体细胞不表达MCP-1。