Expression of gonadotropin-releasing hormone (GnRH) receptor gene is altered by GnRH agonist desensitization in a manner similar to that of gonadotropin beta-subunit genes in normal and castrated rat pituitary.

Abstract

It was previously established that the administration of a potent GnRH agonist such as triptorelin (D-Trp6-GnRH) induced desensitization of pituitary gonadotropic cells, resulting in decreased expression of gonadotropin beta-subunit genes and the suppression of LH and FSH synthesis and release. Binding of GnRH to the pituitary is also affected by agonist treatment. To examine the desensitizing effects of GnRH agonist on the expression of the pituitary GnRH receptor (GnRH-R) gene, male rats were given triptorelin (long-acting formulation, 300 micrograms/kg), and levels of GnRH-R messenger RNA (mRNA) were determined by Northern and dot blot hybridization to a 32P-labeled rat complementary DNA probe. Abundances of gonadotropin alpha-subunit, LH beta, and FSH beta mRNAs were examined in parallel, using appropriate probes. A rapid time-dependent decrease in the level of GnRH-R mRNA was observed in rats after triptorelin administration. A minimum residual level of mRNA, in the range of 20-25% of the initial value, was attained as early as 5 h after treatment. Levels further stabilized to 25-30% after a small transient increase to 45% on day 5. A single injection was effective for at least 30 days, after which GnRH-R mRNA levels slowly returned to normal, suggesting a progressive abolition of agonist effects. A concomitant acute depletion of mRNA levels was observed for LH beta and FSH beta (50% decrease in about 48 and 3 h, respectively), whereas the alpha-subunit message increased (rapidly reaching a level 1.8-fold that in control rats after 1-2 days). Castration induced a 3.8-fold elevation in the amounts of GnRH-R mRNA after 3 weeks, whereas alpha, LH beta, and FSH beta mRNAs increased by 6.2-, 7.9-, and 4.2-fold, respectively, compared to corresponding values in intact animals. Administration of the GnRH agonist readily prevented, for as long as 3 weeks, the stimulatory effects of castration on the GnRH-R mRNA and mRNAs for the beta-subunit of gonadotropins, but not for the alpha mRNA, which remained at a high level. When triptorelin was administered 3 weeks postoperatively, the castration-induced increase in LH beta and FSH beta was totally abolished, and no significant effect was noted on alpha-subunit mRNA. In conclusion, these data demonstrate that expression of the GnRH-R gene is subject to regulation and depends on GnRH stimulation, in a manner that indicates susceptibility to desensitizing action by the long-acting GnRH analog, triptorelin.(ABSTRACT TRUNCATED AT 400 WORDS)

Luteinizing hormone-releasing hormone (LH-RH) antagonist Cetrorelix down-regulates the mRNA expression of pituitary receptors for LH-RH by counteracting the stimulatory effect of endogenous LH-RH

“…It is well known that the elimination of the negative feedback of ovarian steroid hormones on the hypothalamus by ovariectomy causes an increase in the pulsatile LH-RH release from hypothalamic neurons and a marked elevation in the LH-RH concentration in the pituitary portal vessels (36). Accordingly, it was reported that endogenous LH-RH stimulates the synthesis and release of gonadotropins, and increases the number and mRNA expression of its own receptor (35). In keeping with these results, we also found that ovariectomy of rats greatly increased the LH secretion and the LH-RH-R mRNA level in the pituitary.…”

“…Much evidence indicates that the receptor number is regulated at the pretranslational level by LH-RH-R gene expression (26,35), but data on the posttranscriptional regulation by recycling, modification, or degradation have also been reported (12).…”

Gonadotropin-releasing hormone and the control of gonadotrope function

“…Indeed, low frequencies in the range one stimulus every two or four hours appear to preferentially increase FSHβ mRNA levels whereas higher frequencies preferentially stimulate LHβ and α subunit mRNA [13,14]. In vivo, permanent exposure to GnRH leads to a more or less rapid depletion of both FSHβ and LHβ mRNA in a manner that suggests a rapid transcription arrest followed by RNA degradation, while the α-subunit mRNA [15][16][17]. The latter data are consistent with current clinical studies showing that long-term administration of GnRH superagonists induces depletion of LH and FSH but increases free α-subunit levels in the serum of treated patients [18].…”

“…To date, however, much more is known from the first approach. Interestingly, excluding the three gonadotropin subunit genes, GnRH regulates genes essentially associated with the signalling network such as the GnRH receptor itself [17,40], NOS I [41], or PKA and PKC subunit isoforms [42,43] as well as transcription factors such as cFos, cJun or Egr-1. Regulation of the GnRH receptor gene followed a similar extreme dependency on the mode of GnRH stimulation previously revealed for the gonadodotropin β-subunit genes [17,40].…”

“…Interestingly, excluding the three gonadotropin subunit genes, GnRH regulates genes essentially associated with the signalling network such as the GnRH receptor itself [17,40], NOS I [41], or PKA and PKC subunit isoforms [42,43] as well as transcription factors such as cFos, cJun or Egr-1. Regulation of the GnRH receptor gene followed a similar extreme dependency on the mode of GnRH stimulation previously revealed for the gonadodotropin β-subunit genes [17,40]. For both the GnRH receptor and the NOS I genes, evidence exists that the expression is up-regulated at proestrus of the ovarian cycle [31,44,45] suggesting a functional adaptation of the signalling machinery at this important physiological stage.…”