Mouse placental cells secrete immunoreactive growth hormone-releasing factor.
Mouse placental cells secrete immunoreactive growth hormone-releasing factor.
复制标题
小鼠胎盘细胞分泌免疫反应性生长激素释放因子。
DOI：
10.1095/biolreprod51.6.1206
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发表时间：
1994
影响因子：
3.6
通讯作者：
Talamantes,F
中科院分区：
文献类型：
--
作者：
Endo,H;Yamaguchi,M;Farnsworth,R;Thordarson,G;Ogren,L;Alonso,FJ;Sakata,M;Hirota,K;Talamantes,F
这项研究的最初目标是建立一种胎盘细胞培养系统，在该系统中可以在几天的时间内检测小鼠生长激素释放因子(MGHRF)的分泌。为了确定妊娠期间胎盘细胞何时开始表达mGHRF，对妊娠6、9、11、13、15、17和18天采集的胎盘总RNA进行了Northern杂交分析。小鼠生长激素受体基因的表达早在妊娠第11天就可检测到。其稳态水平在第15-17天达到最大值，然后在第18天略有下降。基底带和迷路分散在胶原酶中，细胞在Percoll梯度上分裂。选择两条带进行进一步研究。在5天的培养期间，两者都释放了大量的免疫活性mGHRF。然后检测细胞长期暴露于8-溴-cAMP和升高细胞内cAMP浓度的药物的影响。用0.5 mM的8-溴-cAMP处理细胞，在培养的第二天，培养液中的mGHRF浓度显著降低。用100 ng/ml霍乱毒素或0.1 mM福司可林处理细胞，也能抑制小鼠GHRF的分泌。8-溴-cAMP的作用呈浓度依赖性，0.1 mM为最低活性浓度。8-溴-cAMP处理也降低了细胞内mGHRF mRNA的稳态水平。
The initial objective of this study was to establish a placental cell culture system in which the secretion of mouse growth hormone-releasing factor (mGHRF) could be examined during a several-day period. To determine when during pregnancy placental cells begin to express mGHRF, Northern blot analysis was carried out on total RNA from placentas collected on Days 6, 9, 11, 13, 15, 17, and 18 of pregnancy. Mouse GHRF mRNA could be detected as early as Day 11 of pregnancy. Its steady-state levels increased to maximum values on Days 15-17 and then declined slightly on Day 18. Placentas from Day 12 of pregnancy were selected for cell culture. The basal zone and labyrinth were dispersed in collagenase, and the cells were fractionated on a Percoll gradient. Two bands of cells were selected for further study. Both released significant amounts of immunoreactive mGHRF during a 5-day culture period. Effects of prolonged exposure of the cells to 8-bromo-cAMP and to agents that elevate intracellular cAMP concentration were then examined. Treatment of the cells with 0.5 mM 8-bromo-cAMP resulted in a significant decrease in the mGHRF concentration of the medium by the second day of culture. Mouse GHRF secretion was also inhibited by treatment of the cells with 100 ng/ml cholera toxin or 0.1 mM forskolin. The effect of 8-bromo-cAMP was concentration-dependent, with 0.1 mM being the lowest concentration that was active. 8-Bromo-cAMP treatment also reduced the steady-state level of mGHRF mRNA in the cells.(ABSTRACT TRUNCATED AT 250 WORDS)