Yamaguchi M, Miyake A
Department of Obstetrics and Gynecology, Osaka University Medical School, Japan.
J Endocrinol Invest. 1996 Sep;19(8):492-8. doi: 10.1007/BF03349006.
To determine whether G proteins are involved in the regulation of mouse placental lactogen-I (mPL-I) and/or mPL-II secretion before midpregnancy, mouse placental tissue from day 7 of pregnancy was dispersed with collagenase, cells were fractionated on a percoll gradient, and the purified trophoblast cells were cultured in a serum-free medium with cholera toxin (CTX) or pertussis toxin (PTX) which modulate the activities of distinct G proteins for 5 days. CTX inhibited both mPL-I and mPL-II secretion, but PTX inhibited mPL-I secretion and stimulated mPL-II secretion in a time- and dose-dependent manner. Addition of both CTX and PTX additionally inhibited mPL-I secretion but did not affect mPL-II secretion. 8-Bromo cAMP, which increases intracellular cAMP accumulation, inhibited both mPL-I and mPL-II secretion similarly to CTX. In contrast, H8, an inhibitor of cAMP-dependent protein kinase A, stimulated both mPL-I and mPL-II secretion. Addition of PTX and H8 synergistically stimulated mPL-II secretion. These findings suggest that G proteins play important roles in regulation of mPL-I and mPL-II secretion before midpregnancy.
为了确定G蛋白是否参与妊娠中期前小鼠胎盘催乳素-I(mPL-I)和/或mPL-II分泌的调节,将妊娠第7天的小鼠胎盘组织用胶原酶分散,细胞在 Percoll 梯度上进行分级分离,纯化的滋养层细胞在无血清培养基中用霍乱毒素(CTX)或百日咳毒素(PTX)培养5天,CTX和PTX可调节不同G蛋白的活性。CTX抑制mPL-I和mPL-II的分泌,但PTX以时间和剂量依赖性方式抑制mPL-I分泌并刺激mPL-II分泌。同时添加CTX和PTX可进一步抑制mPL-I分泌,但不影响mPL-II分泌。8-溴环磷酸腺苷(8-Bromo cAMP)可增加细胞内cAMP积累,与CTX类似,它抑制mPL-I和mPL-II的分泌。相反,cAMP依赖性蛋白激酶A的抑制剂H8刺激mPL-I和mPL-II的分泌。添加PTX和H8可协同刺激mPL-II分泌。这些发现表明,G蛋白在妊娠中期前mPL-I和mPL-II分泌的调节中起重要作用。