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At least 235 records · Page 13Linked to original sources

Association of human follitropin (FSH) receptor with splicing variant of human lutropin/choriogonadotropin receptor negatively controls the expression of human FSH receptor.

A splice variant of human lutropin (LH)/choriogonadotropin (CG)-receptor [hLHR(exon 9)] that lacks exon 9 was previously cloned in the corpus luteum of a woman with a normal menstrual cycle. Supported by a detergent-soluble binding assay and a receptor biotinylation experiment, the receptor binding assay shows hLHR(exon 9) is neither expressed at the cell surface nor has the capability of binding to hCG. In addition, hLHR(exon 9) was confirmed in the endoplasmic reticulum (ER) by endoglycosidase H treatment. A coimmunoprecipitation experiment clearly showed that hLHR(exon 9) and constitutively inactivate mutant-LHRs, which stay in the ER, form an association with the human follitropin (FSH)-receptor (hFSHR). This suggests that in the presence of mutant-LHR, hFSHR, which is trapped in the ER and associated with hLHR(exon 9), is unable to come up to the plasma membrane. This phenomenon is specific among gonadotropin receptors because human TSH receptor failed to be coimmunoprecipitated. Furthermore, this receptor complex attenuated the hFSHR receptor protein level within the cells, which impaired cAMP production. To elucidate the mechanism underlying the decrease in hFSHR protein by this receptor complex, we performed a Percoll fractionation experiment, which indicated that the receptor complex drove hFSHR to the lysosome instead of the plasma membrane. These results reveal a novel mechanism of FSHR expression regulation.

Alternative Splicing↗

Pharmacological characterization of soluble human FSH receptor extracellular domain: facilitated secretion by coexpression with FSH.

Follicle-stimulating hormone (FSH) is a member of the glycoprotein hormone family that regulates gametogenesis and steroidogenesis. Glycoprotein hormones signal through a unique class of G-protein-coupled receptors (GPCRs) that have a long extracellular domain (ECD), which is the primary site for hormone binding. The hFSHR-ECD was expressed in insect cells as a C-terminal, epitope-tagged protein resulting in production of soluble active receptor in the intracellular compartment and in the secreted culture medium. Coexpression of hFSHR-ECD with FSHbeta or FSHalpha/beta increased the secretion of the truncated receptor. Pharmacological studies to assess ligand-receptor interactions without the transmembrane domains showed higher affinity values (K(D)S) for [125I]hFSH using mammalian-expressed full-length receptor, secreted hFSHR-ECD, or secreted hFSHR-ECD coexpressed with FSHbeta, whereas the K(D) value for hFSHR-ECD coexpressed with FSHalpha/beta subunits showed lower affinity. Competition of other glycoprotein hormones for secreted hFSHR-ECD coexpressed with FSHbeta or mammalian full-length hFSHR resulted in similar binding profiles, indicating analogous pharmacology. Finally, we have demonstrated that a small molecule, suramin, which has been reported to interact with the mammalian full-length FSHR, competes for the binding of [125I]hFSH by interacting directly at the hFSHR-ECD.

Animals↗

Relationship of follicle stimulating hormone (FSH)-sensitive adenylate cyclase activity to FSH binding in immature rat ovaries following administration of pregnant mare serum gonadotrophin.

The time and dose-response relationships of human follicle stimulating hormone (hFSH)-sensitive adenylate cyclase activity to hFSH binding was studied in the immature rat ovary following an sc injection of pregnant mare serum gonadotrophin (PMSG). When an optimal dose of PMSG (10 IU/rat) was administered, a marked increase in hFSH-sensitive activity was observed at day 2, followed by a sharp decline at day 3. This was accompanied by a parallel rise and fall in ovarian hFSH binding activity. When immature rats were given various doses (5-100 IU/rat) of PMSG for 2 days, hFSH-sensitive adenylate cyclase activity increased sharply and maximal stimulation was obtained at 10 IU/rat. A close correlation was also observed with respect to dose-response for hCG-sensitive adenylate cyclase and hCG binding activities. It is concluded that: 1) PMSG administration with an optimal dose to the immature rat induced ovarian FSH and LH-hCG receptors, and an adenylate cyclase system highly sensitive to hFSH and hCG, and 2) the acquisition and responsiveness of adenylate cyclase to gonadotrophins are closely related to the appearance and the numbers of gonadotrophin receptors.

Adenylyl Cyclases↗

Controlled ovarian stimulation with exclusive FSH followed by stimulation with hCG alone, FSH alone or hMG.

OBJECTIVE: To assess if low-dose hCG is similar to hMG and to rFSH in the late follicular phase. STUDY DESIGN: In a prospective randomized controlled trial, 51 patients undergoing controlled ovarian stimulation received ovarian priming with rFSH and then received hCG (200 IU/day) (hCG group, n=17), hMG (225 IU/day) (hMG group, n=17) or rFSH (200 IU/day) (FSH group, n=17) in the late stage of follicular development. Parameters of follicular response and serum estradiol, progesterone and testosterone levels were assessed. RESULTS: Pre-ovulatory ovarian follicle occurrence and length of treatment were similar among the three treatment groups. Serum progesterone level on the day of pre-ovulatory hCG was significantly higher in the hCG group than in the hMG or rFSH group. Clinical pregnancy rates were similar for all groups. The total cost of treatment was significantly lower for the hCG group than for the groups supplemented with hMG or rFSH. CONCLUSIONS: LH in the form of low-dose hCG during the late follicular phase induced the same follicular pattern as hMG and rFSH after ovulation induction. The procedure using hCG produced pregnancy rates similar to those obtained using hMG and rFSH, even though the patients showed higher serum progesterone levels on the hCG day.

Adult↗

[Comparative analysis of multifollicular development with the application of recombinant FSH vs. urinary FSH in the results of in vitro fertilization].

The interactive function between intrafollicular factors related with ovum competence and embryo development played an important role getting high quality embryos during the embryo transference in in-vitro fertilization programs. Urinary gonadotropins are used widely to get homogeneous follicles during ovarian stimulation. Recombinant technology has been applied in these compounds to correlate with a best egg formation and higher pregnancy rates. We developed a prospective, linear and comparative study to aim the ovum/embryo development using urinary and recombinant FSH. A total of 100 patients were included in the study, 56 patients received rFSH (group I) and 44 hMG (group II). There were not differences in age, weight and body mass index as well as ovarian reserve. However, patients who used urinary gonadotrophins required a higher number of ampoules [31.7 +/- 8.6 vs. 20.7 +/- 6.4 (p < 0.001)]. No differences in peak E2, day of hCG and endometrial thickness. Although, no differences in number of egg mature, the fertilization rate was higher in the recombinant group [5.9 +/- 3.7 vs. 3.4 +/- 2.3 (p < 0.02)]. A higher number of embryo transfer were observed in the same group (3.4 +/- 1.7 vs. 1.9 +/- 2.2 (p < 0.004)]. The pregnancy rates were 34.3% and 29.6% for each group respectively.

Adult↗

Two receptor binding regions of human FSH show sense-antisense similarity to the human FSH receptor.

The sequences of two receptor binding regions of the beta-subunit of the human follicle-stimulating hormone (hFSH-beta) were compared with the DNA-derived antisense peptide sequence of the hFSH receptor. A striking sense-antisense similarity was established between these receptor binding regions and the hFSH receptor. Based on this sense-antisense similarity four putative hormone binding regions on the N-terminal extracellular region of the hFSH receptor are identified.

Amino Acid Sequence↗

Targeted expression of human FSH receptor Asp567Gly mutant mRNA in testis of transgenic mice: role of human FSH receptor promoter.

AIM: To specifically express the Asp567Gly human follicle-stimulating hormone receptor (FSHR) under the control of its promoter to evaluate the phenotypic consequences in the presence of normal pituitary function. METHODS: We produced transgenic mice overexpressing the Asp567Gly human FSHR under the control of a 1.5kb 5'-flanking region fragment of its promoter. RESULTS: Mice were phenotypically normal and fertile. In males, mRNA could be detected in the testis and the brain, indicating that the 1.5kb promoter fragment drives expression not only in the gonads. The testis weight/body weight ratio and the testosterone levels in transgenic and non-transgenic littermates were similar. By in situ hybridisation we found that the transgenic FSHR was highly expressed in Sertoli cells, spermatocytes and round spermatids. However, a radioligand receptor assay failed to show a significant difference in total FSHR binding sites in testis homogenates of transgenic and wild type animals, suggesting that the transgenic FSHR is probably not translated into functional receptor protein. CONCLUSION: A 1.5kb 5'-region of the human FSHR drives mRNA expression of the transgene in the testis but leads to ectopic expression in germ cells and in the brain. No phenotypic consequences could be documented due to the lack of protein expression.

Animals↗

A randomized, multicenter study comparing the efficacy of recombinant FSH vs recombinant FSH with Ganirelix during superovulation/IUI therapy.

OBJECTIVE: The purpose of this study was to determine if use of a Gonadotropin releasing hormone (GnRH) antagonist, Ganirelix (Antagon), can improve pregnancy rates during superovulation with recombinant follicle-stimulating hormone (rFSH) followed by intrauterine insemination (IUI). STUDY DESIGN: This was a multicenter, prospective, randomized, open-label, assessor-blind, controlled trial of females (n = 54), ages 18 to 39 undergoing superovulation/IUI with up to 4 cycles of superovulation/IUI without Ganirelix (n = 66), or up to 4 cycles of superovulation/IUI with the addition of Ganirelix (n = 52). RESULTS: No statistically significant difference in clinical pregnancy rates per cycle initiated was found for patients in the treatment or control group (12% vs 7%, P =.29). Other variables assessed, including endometrial thickness, size of follicles, peak serum estradiol levels, mid-lutea progesterone levels, and total vials of rFSH used also showed no statistically significant difference. CONCLUSION: Superovulation/IUI cycles using Ganirelix produce similar pregnancy rates when compared with cycles not using a GnRH antagonist, although there is a trend towards better pregnancy rates in cycles with Ganirelix.

Adult↗