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PubMed2000

Regulation of endometrial cancer cell growth by luteinizing hormone (LH) and follicle stimulating hormone (FSH).

Gonadotrophin releasing hormone analogues (GnRHa) have been used to treat recurrent endometrial cancer. However, the mode of action is uncertain. Our previous studies showed no direct effect of GnRHa on endometrial cancer cell growth in vitro. We have now examined the effect of luteinizing hormone (LH) and follicle stimulating hormone (FSH) on endometrial cancer cell growth. The aim was to determine whether suppression of pituitary LH and FSH by GnRHa could explain the tumour regression seen in up to 44% of patients treated with this drug. We show that recombinant human LH and FSH (rhLH and rhFSH) produce a concentration dependent stimulation of the endometrial cancer cell line HEC-1A, in serum-free medium (maximum increase of 62 and 50% respectively relative to untreated controls). This increase is equivalent to that obtained by addition of 10% newborn calf serum. Growth of the Ishikawa cell line in culture increases in the presence of rhLH (maximum increase of 67%) but not with rhFSH. Using RT-PCR, we show that the Ishikawa cell line intermittently expresses receptor mRNA of LH but not of FSH; there is no expression of either mRNA by HEC-1A. Classically, both LH and FSH act via cAMP linked membrane receptors. However, neither rhLH nor rhFSH elicit cAMP production in either of our endometrial cancer cell lines. Thus, although a growth response to LH and FSH can be shown, and some cells express the LH receptor, stimulation appears to be via a pathway separate from that of the classical gonadotrophin receptor.

Cell Division↗
PubMed2001

Functional importance of bovine myometrial and vascular LH receptors and cervical FSH receptors.

Bovine myometrium and cervix contain luteinizing hormone/human chorionic gonadotropin (LH/hCG) binding sites, LH receptor (LH-R) messenger RNA (mRNA), and LH-R protein. Expression of LH-R is dependent on the stage of the cycle. LH-R expression is high during the luteal phase but weak during the follicular phase. In both myometrium and cervix, LH activates both the adenylate cyclase and phospholipase C pathways, and the effect of LH on both pathways at each stage of the cycle is correlated with the amount of LH-R present in the tissue. Because activation of cyclic AMP (cAMP) is associated with myometrial quiescence, we suggest that LH activation of uterine cAMP could serve to keep the uterus quiescent during the luteal phase. On the other hand, in the uterine vein LH-R mRNA and LH-R are maximal during preestrus/estrus as opposed to the luteal phase. In the uterine vein, LH increases the expression of cyclooxygenase and production of both prostaglandin E2 (PGE2) and PGF2 alpha. Because PGF2 alpha is the physiological luteolytic signal in the cow, we suggest that this increase in prostaglandin production by the uterine vein is part of the physiological events leading to luteolysis. In addition to uterine LH-R, the bovine cervix at preestrus/estrus has high levels of follicle-stimulating hormone receptor (FSH-R) and its corresponding mRNA. As with LH-R, activation of FSH-R by FSH is associated with activation of a G protein-coupled receptor family that mediates the cAMP and inositol phosphate signaling pathways. Activation of these signaling pathways is associated with an increase in the expression of cyclooxygenase and production of PGE2. Because expression of the FSH receptor was maximal at the time of the FSH peak in the blood, we suggest a physiological role for FSH in the cervix relaxation and opening at estrus.

Adenylyl Cyclases↗
PubMed2004

Human cumulus-free oocyte maturational profile and in vitro developmental potential after stimulation with recombinant versus urinary FSH.

BACKGROUND: This study compares the influence of recombinant (r)FSH and urinary (u)FSH stimulation on oocyte and embryo quality in patients undergoing ICSI. METHODS AND RESULTS: Denuded oocyte maturity in ICSI patients was graded on a scale from metaphase II (MII) to prophase for nuclear maturation of oocytes. The relationships of cumulus-free oocyte maturational profiles with in vitro outcome parameters were evaluated. In the study population with an unknown distribution of FSH receptor polymorphisms, the ovarian response to rFSH stimulation was significantly different from that of uFSH stimulation, including lower number of oocytes retrieved/oocytes in MII, higher fertilization rates and higher good quality embryo rates. In the study population with a similar distribution of FSH receptor polymorphisms, the ovarian responses to rFSH were lower numbers of oocytes in MII, higher fertilization rates and higher good quality embryo rates, but the total number of oocytes retrieved was not influenced, in comparison with ovarian stimulation with uFSH. CONCLUSIONS: rFSH stimulation appears to influence oocyte quality and subsequent embryo quality in comparison with uFSH stimulation. FSH receptor polymorphisms seem to be an intrinsic factor influencing the ovarian response to FSH stimulation.

Culture Techniques↗
PubMed1996

Ontogenesis of LH and FSH receptors in postnatal rabbit testes: age-dependent differential expression of long and short RNA transcripts.

The ontogeny of testicular LH and FSH receptors was studied in New Zealand rabbits from 20 to 180 days postpartum. The concentrations of free receptors (per mg total proteins) were very low at day 20. They increased steeply at day 30 for the LH receptor and at day 50 for the FSH receptor. Three RNA bands (1.2, 2.5 and 3 kb) were repeatedly detected on northern blots for the LH receptor and two bands (1.2 and 2.2 kb) were detected for the FSH receptor. The 1.2 kb band (which cannot give rise to full-length, membrane-anchored receptor) was present throughout the 20-180 day period for each receptor. However, the higher molecular mass bands were nearly undetectable at day 20. The 2.5 and 3 kb bands of the LH receptor increased twofold between day 20 and day 120, while the 2.2 kb band of the FSH receptor increased fivefold between day 20 and day 75. Thus the very low concentrations, or even absence, of the larger transcripts of both LH and FSH receptors were correlated with the inability to detect their cognate protein until 20 days of age. Subsequently, coordinated increases in high molecular mass transcripts and protein were observed for both receptors. Total LH receptor content increased in parallel to the previously reported increase in plasma testosterone between day 65 and day 100. FSH receptor density began to increase steeply at day 50, just at the onset of spermatogenesis. Thus, postnatal testicular development in the rabbit seems to entail the transcription of high molecular mass, translatable transcripts of the gonadotrophin receptors.

Animals↗
PubMed1990

Available FSH receptors in adult rat testis in vivo.

It has been suggested that the tight junctions formed between Sertoli cells during the peripubertal period constitute a barrier to circulating FSH in the adult testis, limiting its access to the lumen of the seminiferous tubules. There are also reports of FSH receptor-binding inhibitors. These observations prompted us to study the extent of FSH receptor availability in vivo in the adult rat. Experimental rats were given an intracardiac injection of rat FSH (rFSH), and the occupied receptor was measured by radioimmunoassay of acid-released rFSH from the testis. In the saline-injected control animals, there were 247 fmol occupied FSH receptors/g testis, and as much as 1788 fmol of the unoccupied high-affinity receptors/g testis, as measured by in-vitro binding studies. After intracardiac injection of increasing amounts of rFSH (up to 606 pmol), receptor occupancy increased to a maximum plateau of only 448 fmol/g testis. In contrast, when rFSH was given by intratesticular injection in order to achieve pharmacological doses in the testis, the maximum binding was 662 fmol/g testis. Scatchard analysis of the in-vivo data revealed, however, that the maximum concentration of the high-affinity receptor was 452 fmol/g testis, a value concordant with the highest in-vivo binding observed in animals given intracardiac rFSH (448 fmol/g). A single injection of the hormone did not induce down-regulation of FSH receptors, regardless of the dose, whereas multiple injections of menotrophin were effective, at least to some extent. Despite the receptor loss, the immediate receptor availability was maintained, suggesting the presence of a receptor pool.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

[Effects of activin and follistatin on FSH receptor mRNA expression of cultured Shao duck granulosa cells].

The aim of this study was to investigate the action of activin and follistatin either alone or in combination on FSH receptor mRNA expression. The results showed that activin alone increased FSH receptor mRNA level in the presence or absence of FSH in cultured granulosa cells, and the stimulating effect of activin on FSH receptor level was dose-dependent. This effect of activin was inhibited by FSP treatment which alone had no effect on FSH receptor expression. From the results, it can be concluded that activin and follistatin both exert autocrine actions on granulosa cells, and the two factors, possibly by regulating FSH receptor expression, may play important roles in follicular development.

Activins↗
PubMedDate not supplied

Selective effects of charge on G protein activation by FSH-receptor residues 551-555 and 650-653.

Two cytosolic regions of the rat testicular FSH receptor (FSHR), residues 533-555 and 645-653, have been identified as G protein-coupling domains. We localized the activity in these domains to their C-terminal sequences, residues 551-555 (KIAKR, net charge +3) and 650-653 (RKSH, net charge +3), and examined the effects of charge on G protein activation by the C-terminal peptides, using synthetic analogs containing additions, through alanine (A) linkages, of arginine (R, +), histidine (H, +) or both. RA-KIAKR (net charge +4) mimicked the effect of FSHR-(551-555) on guanine nucleotide exchange in rat testis membranes, but reduced its ability to inhibit FSH-stimulated estradiol biosynthesis in cultured rat Sertoli cells. Further increasing net charge by the addition of H (HARA-KIAKR, net charge +5) increased guanosine 5'-triphosphate (GTP) binding, but eliminated FSHR-(551-555) effects on FSH-stimulated steroidogenesis. HA-RKSH (net charge +4) significantly inhibited guanine nucleotide exchange in rat testis membranes, but stimulated basal and potentiated FSH-induced estradiol biosynthesis in cultured rat Sertoli cells. Addition of two H residues (HAHA-RKSH, net charge +5) restored GTP binding and further potentiated basal and FSH-stimulated steroidogenesis. These results suggest that positive charges in G protein-coupling domains of the FSHR play a role in modulating G protein activation and postbinding effects of FSH, such as steroidogenesis.

Amino Acid Sequence↗
PubMed1997

Increased LH and FSH secretion after cranial irradiation in boys.

The effect of high-dose cranial- and craniospinal irradiation and chemotherapy on the gonadotropin-sex steroid axis was studied during different stages of puberty by measuring pulsatile secretion of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone. The patients were thirteen boys who had been treated for malignant brain tumor residing well away from the hypothalamo-pituitary region. The median time to follow-up was 9 (1-16) years. The onset of puberty was early in the patients, median 10.5 years, compared to the average age for Swedish boys, which is at median 12.4 years. There was, before puberty, no significant difference in LH and FSH secretion between patients and a control group of normal boys. In early, mid- and late stages of puberty, however, LH and FSH secretion was increased in the patients overall, whereas testosterone secretion was maintained within the normal range in spite of signs of gonadotoxocity with small testicular volumes. These results indicate that the vulnerable parts of the gonadotropin releasing hormone (GnRH)-gonadotropin (LH, FSH)-gonadal axis are the regulatory system that determines the timing of pubertal induction and the gonads. The GnRH-LH, FSH-releasing neurons appear relatively resistant to cranial irradiation as they are able to respond with supranormal LH and FSH levels for long periods of time after treatment.

Adolescent↗
PubMed1999

Effect of season and photoperiod on FSH in male great tits.

The present study investigated (1) the photoperiodic threshold for an increase in FSH and LH secretion in male great tits (Parus major) from south Sweden and (2) seasonal changes in FSH secretion patterns in short day (SD; 8 L:16 D) and long day (LD; 20 L:4 D) exposed great tits. When fully photosensitive great tits were exposed to a simulated vernal increase in day lengths plasma levels of FSH started to rise at a shorter day length (10.5 h) than did LH (11 h). Once the photoperiodic threshold was exceeded FSH levels increased slowly, but continuously, and reached maximum levels when day lengths had reached 13-14 h, whereas LH levels rose to maximum levels more or less immediately. Great tits break photorefractoriness in September. Exposing sham-operated and castrated great tits to SD or LD in October and February resulted in clear seasonal differences in their FSH cycles. At both times the great tits responded to castration and exposure to long days. However, castration and photoperiodic FSH responses were not only more rapid in February, but also of a higher magnitude.

Animals↗
PubMed1999

Ontogenetic profile of FSH and LH in Rana esculenta.

Circulating levels and pituitary content of FSH and LH were determined by specific radioimmunoassays in Rana esculenta starting a few days after hatching until the completion of metamorphosis. Both gonadotropins were found in the pituitary as well as in the blood plasma at all stages of development examined here. The plasma concentrations of FSH and LH were more or less uniform during pre- and prometamorphosis, but increased significantly at the onset of metamorphic climax. The plasma levels of FSH and LH remained high at the completion of metamorphosis. The pituitary content of FSH and LH was low in early premetamorphosis. It increased slightly through prometamorphosis and metamorphic climax, following which a highly significant increase occurred. Whereas plasma concentrations of FSH and LH were essentially similar within a single stage of development, the pituitary FSH content was severalfold higher than pituitary LH. The significance of these results is discussed in relation to the functional maturation of the brain-pituitary-gonadal axis in the frog.

Animals↗
PubMed1999

Influence of corticosterone on FSH-induced ovarian recrudescence in the lizard Mabuya carinata.

Administration of bovine FSH (10 IU/lizard/alternate day for 30 days) in the postbreeding quiescent phase of the ovarian cycle caused a significant increase in the mean number of oogonia and oocytes, the relative weight of the oviduct, and the liver and serum estradiol levels compared to those of controls. In addition, the FSH-treated lizards showed a vitellogenic growth of follicles and development through to preovulatory follicles. However, the administration of corticosterone simultaneously with FSH (10 IU FSH + 40 microgram corticosterone/lizard/alternate day for 30 days) did not result in these changes and the ovaries resembled those of controls. The results indicate the absence of ovarian refractoriness to gonadotropic stimulation during the quiescent phase of the reproductive cycle and inhibition of FSH-induced ovarian recrudescence by corticosterone. It is suggested that corticosterone treatment reduces FSH-induced steroidogenic activity of the ovary, leads to impairment in vitellogenin secretion by the liver, and results as well in the failure of vitellogenic follicular growth in Mabuya carinata.

Animals↗
PubMed2000

Spatio-temporal expression of TSH beta and FSH beta genes in normally metamorphosing, metamorphosed, and metamorphosis-arrested Hynobius retardatus.

Expression patterns of TSH beta and FSH beta genes were investigated in normally metamorphosing and metamorphosed individuals and in goitrogen-treated (TU-SPC), metamorphosis-arrested larvae of a Japanese salamander, Hynobius retardatus, a particular population of which had been reported to show neoteny. Cloned sequences for Hynobius TSH beta and FSH beta covered 66 and 56% of full-length genes, respectively, and both showed relatively high similarity to those of other vertebrates. TSH beta-expressing cells were observed at the ventro-caudal region of the pars distalis of controls, whereas an extraordinarily large number of cells which occupied the whole area of the pituitary glands strongly expressed TSH beta in TU-SPCs. In contrast, no FSH beta-expressing cells were observed in the pituitary glands of controls, whereas several positive cells were observed in the pituitary glands of TU-SPCs. Northern blot analyses revealed a rapid increase of TSH beta expression in the pituitary gland of TU-SPCs that finally reached the level of 20- to 30-fold as much as that of controls. Surprisingly, the expression of FSH beta also increased in the pituitary gland of TU-SPCs compared with that of controls. On the other hand, triiodothyronine administration to the control juveniles reduced the expression of TSH beta compared with saline-administrated controls, suggesting that thyroid hormone (TH) exerts a repressing effect on TSH beta gene. These results suggest that TSH beta overexpression in TU-SPCs must be caused by a deprivation of TH which plays a role in the negative feedback system and that FSH beta expression also might be enhanced by mechanisms similar to those of TSH beta, probably due to certain overlapping of the endocrinological regulation of TSH and FSH in H. retardatus.

Amino Acid Sequence↗
PubMed2000

Corticosterone inhibits normal and FSH-induced testicular recrudescence in the lizard, Mabuya carinata.

Administration (ip) of 1, 10, or 20 microg corticosterone (alternate days for 30 days) to adult male Mabuya carinata did not affect the seasonal recrudescence of spermatogenesis whereas administration of 40 microg corticosterone did result in inhibition of spermatogenesis. Further, administration of FSH (10 IU/lizard/alternate day for 30 days) during the quiescent phase of the testicular cycle stimulated spermatogenetic and steroidogenic activity of the testis as shown by significant increases in the mean number of spermatogonia, spermatocytes, and spermatids and serum levels of testosterone. In addition there were abundant spermatozoa in the lumen of the tubules in FSH-treated lizards. Administration of 10 IU FSH + 40 microg corticosterone (per lizard on alternate days for 30 days) increased the mean number of primary and secondary spermatocytes whereas the mean number of spermatids did not show significant variation compared with that of controls. Further, the mean numbers of spermatocytes and spermatids and serum levels of testosterone were significantly less when compared to those of FSH alone treated lizards. In addition, FSH-induced development of epididymis was also inhibited by corticosterone treatment. The results indicate that corticosterone inhibits FSH-induced testicular recrudescence, possibly by suppressing testosterone secretion in M. carinata.

Animals↗
PubMed1975

Synthesis of estradiol-17 beta by Sertoli cells in culture: stimulation by FSH and dibutyryl cyclic AMP.

Sertoli cells isolated from the testes of 18 to 20-day old rats synthesize estradiol-17 beta when grown in primary culture in the presence of testosterone (5 X 10(-7) M). After an initial lag phase of about 2 hours, follicle-stimulating hormone (FSH) stimulates synthesis of estradiol up to 50-fold, synthesis being approximately linear for 24 hours when medium is changed every 2-6 hours. Luteinizing hormone (LH) causes only a marginal stimulation, at concentrations consistent with contamination with FSH. Dibutyryl cyclic 3',5'-adenosine monophosphate (dbcAMP) (10(-4)M) mimicks the effect of FSH. No significant synthesis occurs in the absence of steroid substrate or in the presence of pregnenolone (5 X 10(-7) M). When cells are pre-incubated for 24 hours with FSH in the absence of testosterone, addition of testosterone results in an immediated increase in estradiol synthesis without a lag phase, suggesting that FSH or dbcAMP are capable of inducing the conditions necessary for stimulation in the absence of testosterone. These observations provide the first direct evidence of estradiol-17 beta synthesis by Sertoli cells from normal animals, and offer evidence that synthesis of this steroid is regulated at the level of the aromatizing enzyme system by FSH and cyclic AMP.

Animals↗
PubMed1984

Ultrastructural immunocytochemical localization of LH and FSH in the pituitary of the untreated male rat.

Rapid freeze-substitution fixation was employed in immunocytochemical studies on the localization of LH and FSH in the typical gonadotrophs of the anterior pituitary in the untreated male rat; a modification of a recently described ferritin antibody method (Inoue et al. 1982) was used in these studies. It was shown that rapid freeze-substitution fixation provides good preservation not only of the ultrastructure but also of the antigenicity. Both LH and FSH were clearly demonstrated in the same gonadotrophic cells, but the subcellular localization of these gonadotrophins differed: (i) LH was mainly located in small secretory granules, 250-300 nm in diameter; (ii) FSH was mainly present in large secretory granules, up to 500 nm in diameter. In the pituitary gland of the adult male rat, all gonadotrophs that react to antibodies against gonadotrophins are characterized by small and large secretory granules. Other types of cells of the anterior pituitary containing either small secretory granules or resembling corticotrophs with secretory granules assembled at cell periphery did not react to either anti-LH beta or anti-FSH beta serum. For light microscopy, the peroxidase antibody method was used. All of the gonadotrophin-positive cells contain both LH and FSH. None of the pituitary cells reacted to antibody against only one gonadotrophin. However, some cells are "LH-rich" while other cells are "FSH-rich".

Animals↗
PubMed1984

The effect of 2 mg estradiol-17 beta plus 1 mg estriol, sequentially combined with 1 mg norethisteroneacetate, on LH, FSH, estradiol-17 beta, progesterone, testosterone and prolactin after ovariectomy.

The object of the study was to see whether maintenance of serum estradiol levels corresponding to the early and mid-follicular phase can prevent the gonadotrophin increase following ovariectomy. We also wanted to study the effect on LH and FSH of an additional dose of 1 mg dose of 1 mg norethisterone acetate administered for 10 days during each month. In 22 women with normal cycles 1 mg of estradiol benzoate was injected i.m. at the time of ovariectomy. From the first post-operative day onwards they received daily doses of 2 mg estradiol and 1 mg estriol in the form of micronized tablets. From the 41st to the 50th day and again from the 69th to the 78th day the patients received additional daily doses of 1 mg norethisterone acetate. LH, FSH, estradiol-17 beta, (E2) progesterone (P), testosterone (T), and prolactin (PRL) were measured in intervals of 2-17 days. Even though the estradiol mean values remained constant in the range of 65-115 pg throughout the period under observation, the LH mean levels increased continuously from 8 to a maximum of 23.9 mU/ml, and the FSH mean level from a pre-operative value of 6-48.0 mU/ml on the 85th day. On the 7th day after the last administration of norethisterone acetate LH was slightly depressed while FSH continued to rise slightly. Both FSH and LH are negatively correlated with E2 and this inverse correlation becomes even more pronounced the more time has elapsed after surgery. These findings suggest that not only the estrogens inhibit FSH and LH but also other steroids and/or nonsteroidal ovarian inhibiting factors.

Adult↗
PubMed1995

Regulation of IGF binding proteins by FSH in human luteinizing granulosa cells.

OBJECTIVE: To evaluate the regulation of the secretion of insulin-like growth factor (IGF) binding proteins (IGFBPs) by FSH in human luteinizing granulosa cells. DESIGN AND RESULTS: Luteinizing granulosa cells were incubated with and without FSH. Levels of IGFBP-1 and -3 in the medium were measured by EIA and RIA, respectively and binding activities of these IGFBP were evaluated by Western ligand blot. FSH inhibited the secretion of IGFBP-1 dose dependently. FSH did not inhibit the secretion of immunoreactive IGFBP-3, but inhibited the binding activity of IGFBP-3. To assess the protease activity, 125I-IGFBP-3 was incubated with the cultured medium. 125I-IGFBP-3 degraded into small fragments when it was incubated with the cultured medium treated with FSH. CONCLUSIONS: FSH enhances the action of IGF-I in human granulosa cells by inhibiting the secretion of IGFBP-1 and the binding activity of IGFBP-3 by stimulating the proteolysis of IGFBP-3.

Blotting, Western↗
PubMedDate not supplied

Cabergoline modulation of alpha-subunits and FSH secretion in a gonadotroph adenoma.

Most non-functioning pituitary adenomas respond poorly to medical therapy. We describe the case of a 62-year-old man who presented with clinical features of an invasive macroadenoma. Baseline hormonal evaluation revealed increased FSH and alpha-subunit (alpha-SU) levels. Transsphenoidal exeresis followed by radiotherapy (RT) was performed. Almost all neoplastic cells were intensely immunoreactive for alpha-SU. On PCR analysis, specific amplification products were observed for somatostatin 2, 3 and 5 receptors as well as for both short and long isoforms of the dopamine D2 receptor. In vitro, alpha-SU and FSH were released into the medium by adenoma cells and increased after TRH stimulation. After surgery, alpha-SU and FSH levels were still elevated. Short-term slow-release lanreotide treatment did not modify either alpha-SU or FSH levels. Cabergoline was started and a fast and long-lasting decrease in alpha-SU and, to a lesser extent, in FSH was observed. The tumor remnant was unmodified on magnetic resonance imaging 3 years after surgery and RT. This case report shows that the in vitro expression of somatostatin receptors may not be directly associated to the in vivo response of alpha-SU and FSH to lanreotide, probably because of a functional uncoupling of the receptors. Cabergoline should be considered as an effective therapy for hormonal, and perhaps proliferative, control of gonadotroph adenoma remnants before the effects of RT are fully effective.

Adenoma↗