Homologous upregulation of GnRH receptor mRNA by continuous GnRH in cultured rat pituitary cells

The present study examined the effects of continuous treatment with gonadotropi-releasing hormone (GnRH) on GnRH receptor (GnRH-R) mRNA levels in dispersed cultures of rat pituitary cells. Pituitary GnRH-R mRNA levels were determined by competitive reverse transcriptase polymerase chain reaction. When pituitary cells were continuously exposed to a low dose of GnRH (0.2 nM), GnRH-R mRNA levels were transiently increased. The levels of GnRH-R mRNA were significantly increased up to 6 h and diminished to untreated levels by 24 h. Luteinizing hormone (LH) release was also increased significantly up to 12 h, maintaining similar levels in LH release therafter. When GnRH antagonist([d-pGlu1, d-Phe2, d-Trp3.6]-LH-RH) was added to the cultures together with GnRH (0.2 nM) for 6 h, the stimulatory effect of GnRH on GnRH-R mRNA levels and LH release was significantly diminished in a dose-related manner. In another experiment, pituitary cells were treated with various doses of GnRH (0.02–200 nM) for a relatively short (6 h) or a longer (24 h) period. When pituitary cells were exposed for 6 h, all doses of GnRH 0.02–200 nM) significantly increased GnRH-R mRNA levels in a dose-dependent manner. By contrast, continuous exposure to GnRH for 24 h was ineffective in changing pituitary GnRH-R mRNA levels at any given doses. These results indicate that the duration of GnRH treatment is critical for upregulation of GnRH-R mRNA by continuous GnRH. When pituitary cells were treated for 6 h with either a continuous mode of GnRH (0.2 nM) or an hourly pulsatile mode of GnRH (0.2 nM, 6 mi/h), both treatment significantly augmented GnRH-R mRNA levels. Thus, the modes of GnRH application, if treated for a relatively short period, do not appear to make a significant difference in upregulation of GnRH-R mRNA levels. Collectively, our data provide strong evidence that continuous GnRH application is able to upregulate pituitary GnRH-R mRNA levels, if treated for a relatively short period (6 h).