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The effect of HGF on LH-dependent androgen production by TIC. A, TIC were incubated in a total volume of 200 l/well with LH (0.003-3.0 ng/ml) in the presence and absence of HGF (50 ng/ml). Control groups received M5a without hormones. B, Dose-dependent effects of HGF on LH-dependent androsterone production by TIC. TIC were treated with LH (0.3 ng/ml), HGF (0.1-100 ng/ml), or LH in the presence of HGF. Cultures in A and B were terminated at 48 h. C, Detailed time course for HGF effects on TIC androgen production. TIC were given LH (0.3 ng/ml), HGF (50 ng/ml), or LH and HGF for 6-48 h. Conditioned media were assayed for androsterone (A, B, and C) and androstenedione (inset) concentration by RIA. Data are the mean SEM of three independent experiments, with three replicates per experiment. Bars with different letters are significantly different (P 0.05).
Source publication
During ovarian follicle growth, precise regulation of the onset of androgen production by ovarian theca-interstitial cells (TIC) is necessary for maintaining follicle viability. Thus, temporary suppression of TIC androgen production in preantral follicles is the key to promoting follicle development. Evidence indicates that this process is coordina...
Contexts in source publication
Context 1
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 2
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 3
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 4
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 5
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 6
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 7
... TIC cultures, LH stimulated a dose-dependent increase in androsterone synthesis compared to controls (Fig. 2a). At the maximal stimulatory concentration of LH (0.3 ng/ml), androstenedione production was also stimulated over that of controls (Fig. 2a, inset). HGF (50 ng/ml) did not alter basal androsterone (Fig. 2a) or androstenedione (Fig. 2a, inset) levels. At the lower concentrations of LH (0.003-0.03 ng/ml), HGF did not affect LH stimulation of androsterone produc- tion. However, at 0.1 ng LH/ml and greater, HGF inhibited androsterone production. At the maximal stimulatory con- centration of LH (0.3 ng/ml), HGF suppressed LH-depen- dent TIC androsterone production by more than 50% (Fig. 2a). Moreover, the suppressive effects of HGF on LH-de- pendent androsterone production were concentration de- pendent (IC 50 1.5 0.01 ng HGF/ml; Fig. 2b). To deter- mine whether HGF selectively impaired the synthesis of androsterone (by interfering with 4-ene-5-reductase and/or 3HSD), or inhibited androgen production at the level of P450 17 , androstenedione levels were also measured. Results showed that LH-stimulated androstenedione production was impaired by HGF (Fig. 2a, inset), and this indicated generalized inhibition of LH-stimulated androgen ...
Context 8
... detailed time-course study showed that at 12-24 h, HGF did not impair LH-dependent androsterone production (Fig. 2c), whereas beginning at 36 h (Fig. 2c) and beyond LH- stimulated androsterone levels were suppressed by HGF. Therefore, it appears that HGF did not completely block TIC differentiation to the androgenic phenotype, and this is sup- ported by the fact that in the presence of HGF, LH-dependent androstenedione and androsterone production remained significantly above control ...
Context 9
... detailed time-course study showed that at 12-24 h, HGF did not impair LH-dependent androsterone production (Fig. 2c), whereas beginning at 36 h (Fig. 2c) and beyond LH- stimulated androsterone levels were suppressed by HGF. Therefore, it appears that HGF did not completely block TIC differentiation to the androgenic phenotype, and this is sup- ported by the fact that in the presence of HGF, LH-dependent androstenedione and androsterone production remained significantly above control ...
Citations
... HGF has also been demonstrated to be an angiogenic factor complementary to FGF2 and VEGF [61]. Previous studies have shown that HGF is an important paracrine/autocrine regulator of theca cell and GC growth and steroidogenesis in ovaries [62,63]. HGF regulates numerous key functions in ovaries, including cell growth, steroidogenesis, and apoptosis of GCs and/or theca cells, which collectively regulate follicular growth and differentiation [48]. ...
Background
Chemotherapy can induce premature ovarian insufficiency (POI) and reduce fertility in young female patients. Currently, there is no effective therapy for POI. Human amnion-derived mesenchymal stem cells (hAD-MSCs) may be a promising seed cell for regenerative medicine. This study investigated the effects and mechanisms of hAD-MSC transplantation on chemotherapy-induced POI in rats.
Methods
Chemotherapy-induced POI rat models were established by intraperitoneal injection of cyclophosphamide. Seventy-two female SD rats were randomly divided into control, POI, and hAD-MSC-treated groups. hAD-MSCs were labeled with PKH26 and injected into the tail veins of POI rats. To examine the underlying mechanisms, the differentiation of transplanted hAD-MSCs in the POI ovaries was analyzed by immunofluorescent staining. The in vitro expression of growth factors secreted by hAD-MSCs in hAD-MSC-conditioned media (hAD-MSC-CM) was analyzed by ELISA. Sixty female SD rats were divided into control, POI, and hAD-MSC-CM-treated groups, and hAD-MSC-CM was injected into the bilateral ovaries of POI rats. After hAD-MSC transplantation or hAD-MSC-CM injection, serum sex hormone levels, estrous cycles, ovarian pathological changes, follicle counts, granulosa cell (GC) apoptosis, and Bcl-2, Bax, and VEGF expression in ovaries were examined.
Results
PKH26-labeled hAD-MSCs mainly homed to ovaries after transplantation.
hAD-MSC transplantation reduced ovarian injury and improved ovarian function in rats with POI. Transplanted hAD-MSCs were only located in the interstitium of ovaries, rather than in follicles, and did not express the typical markers of oocytes and GCs, which are ZP3 and FSHR, respectively. hAD-MSCs secreted FGF2, IGF-1, HGF, and VEGF, and those growth factors were detected in the hAD-MSC-CM. hAD-MSC-CM injection improved the local microenvironment of POI ovaries, leading to a decrease in Bax expression and an increase in Bcl-2 and endogenous VEGF expression in ovarian cells, which inhibited chemotherapy-induced GC apoptosis, promoted angiogenesis and regulated follicular development, thus partly reducing ovarian injury and improving ovarian function in rats with POI.
Conclusions
hAD-MSC transplantation can improve ovarian function in rats with chemotherapy-induced POI at least partly through a paracrine mechanism. The presence of a paracrine mechanism accounting for hAD-MSC-mediated recovery of ovarian function might be attributed to the growth factors secreted by hAD-MSCs.
... Beyond that, VEGF is a powerful survival factor for ovarian granulosa cell apoptosis and ovarian follicular atresia [42,43]. Apart from VEGF, HGF is an important element of the internal follicular environment that accelerates the viability of growing follicles and enhances the proliferation of ovarian surface epithelium in order to replenish the area damaged due to expulsion of the ovum during ovulation [44,45]. HGF, expressed both in thecal cells and granulosa cells of rat ovaries, may play its function as a modulator of the mesenchymal-epithelial cell reciprocities between theca cells and granulosa by facilitating cell proliferation and steroid hormone production [46]. ...
Background
Human umbilical cord mesenchymal stem cells (hUCMSCs) are a type of pluripotent stem cell which are isolated from the umbilical cord of newborns. hUCMSCs have great therapeutic potential. We designed this experimental study in order to investigate whether the transplantation of hUCMSCs can improve the ovarian reserve function of perimenopausal rats and delay ovarian senescence. Method
We selected naturally aging rats confirmed by vaginal smears as models of perimenopausal rats, divided into the control group and the treatment group, and selected young fertile female rats as normal controls. hUCMSCs were transplanted into rats of the treatment group through tail veins. Enzyme-linked immunosorbent assay (ELISA) detected serum levels of sex hormones, H&E staining showed ovarian tissue structure and allowed follicle counting, immunohistochemistry and western blot analysis revealed ovarian expression of hepatocyte growth factor (HGF), vascular endothelial cell growth factor (VEGF), and insulin-like growth factor-1 (IGF-1), polymerase chain reaction (PCR) and western blot analysis revealed hUCMSCs expression of HGF, VEGF, and IGF-1. ResultsAt time points of 14, 21, and 28 days after hUCMSCs transplantation, estradiol (E2) and anti-Müllerian hormone (AMH) increased while follicle-stimulating hormone (FSH) decreased; ovarian structure improved and follicle number increased; ovarian expression of HGF, VEGF, and IGF-1 protein elevated significantly. Meanwhile, PCR and western blot analysis indicated hUCMSCs have the capacity of secreting HGF, VEGF, and IGF-1 cytokines. Conclusions
Our results suggest that hUCMSCs can promote ovarian expression of HGF, VEGF, and IGF-1 through secreting those cytokines, resulting in improving ovarian reserve function and withstanding ovarian senescence.
... It was suggested by Zachow and Uzumcu in 2007 (30), that there is an intriguing parallelism between male and female gonad that hypothesizes some endocrine implications of HGF in the physiology of both organs. It has been reported, also, that HGF can down-modulate ovarian steroidogenesis suppressing FSHdependent 17β-estradiol production by directly impairing CYP19 enzyme (60,61). It is well known that Sertoli cells represent the testicular counterpart of granulosa cells and are the testicular source of 17β-estradiol. ...
... Intriguingly, the steroid modulator activity is not confined in male gonad but was also described in theca cells, which are the Leydig cell ovarian counterparts. In rat theca cells, in fact, HGF suppressed LH-dependent androsterone secretion, while stimulated basal and LH-induced progesterone production (60). It is fair to highlight the parallelism between male and female gonad and to notice that steroid production modulation in response to local cues is quite relevant for gonad physiology since sex hormones are not only important for endocrine homeostasis via pituitary-gonadal axis cross-talk but also act as paracrine regulators of male and female gametogenesis. ...
In the last decades, a growing body of evidence has been reported concerning the expression and functional role of hepatocyte growth factor (HGF) on different aspects of testicular physiology. This review has the aim to summarize what is currently known regarding this topic. From early embryonic development to adult age, HGF and its receptor c-Met appeared to be clearly detectable in the testis. These molecules acquire different distribution patterns and roles depending on the developmental stage or the post-natal age considered. HGF acts as a paracrine modulator of testicular functions promoting the epithelium–mesenchyme cross-talk as described even in other organs. Interestingly, it has been reported that testicular HGF acts even as an autocrine factor and that its receptor might be modulated by endocrine signals that change at puberty: HGF receptor expressed by Sertoli cells, in fact, is up-regulated by FSH administration. HGF is in turn able to modify endocrine state of the organism being able to increase testosterone secretion of both fetal and adult Leydig cells. Moreover, c-Met is expressed in mitotic and meiotic male germ cells as well as in spermatozoa. The distribution pattern of c-Met on sperm cell membrane changes in the caput and cauda epididymal sperms and HGF is able to maintain epididymal sperm motility in vitro suggesting a physiological role of this growth factor in the acquisition of sperm motility. Noteworthy changes in HGF concentration in seminal plasma have been reported in different andrological diseases. All together these data indicate that HGF has a role in the control of spermatogenesis and sperm quality either directly, acting on male germ cells, or indirectly acting on tubular and interstitial somatic cells of the testis.
... When combined with Stem Cell Factor/kit ligand (SCF), IGF-1 increased mRNAs of steroid acute regulatory protein (StAR), CYP11A1, CYP17, 3β-hydroxysteroid dehydrogenase and LH receptor (23). Although expressed by both GCs and TCs, hepatocyte growth factor reduced CYP17 expression and androgen secretion from rat TCs in the presence of LH (24). Overall it appears that there is a complex network of signals between GCs and TCs that act to finetune LH-and FSH-regulation of ovarian steroid production. ...
Objective:
To test whether and to what extent inhibin mediates Cyp17 messenger RNA (mRNA) expression in theca cells (TCs) in response to FSH stimulation of granulosa cells (GCs).
Design:
Ex vivo and in vitro experimental study.
Setting:
University.
Animal(s):
Immature female Sprague Dawley rats.
Intervention(s):
Ovarian tissue explants and isolated theca cell preparations with or without GCs were treated with FSH, inhibin, inhibin antibody, or β-glycan antibody.
Main outcome measure(s):
As a key enzyme in androgen production, Cyp17 mRNA levels were measured by real-time reverse transcription-polymerase chain reaction.
Result(s):
After 24 hours, Cyp17 mRNA expression was dose-dependently increased by FSH in ovarian tissue explants and theca cells, suggesting that paracrine factor(s) secreted from GCs in response to FSH mediates Cyp17 mRNA expression in TCs. Antibodies against inhibin and inhibin coreceptor, β-glycan, blocked the stimulatory effect of FSH on Cyp17 mRNA expression. However, inhibin alone did not increase Cyp17 mRNA level to the same extent.
Conclusion(s):
These findings suggest a role for inhibin in the paracrine regulation of TC Cyp17 mRNA expression by GCs influenced by FSH; however, other paracrine factors produced by GCs by virtue of FSH seem to be required.
... HGF was reported as a growth factor that controls several key functions, including the regulation of growth and differentiation of ovarian follicles. It has been reported that the HGF/ c-Met signaling modulates every major component of folliculogenesis, including steroidogenesis, the growth of theca and granulosa cells, and apoptosis of granulosa cells (22,6). Nevertheless, it is currently unknown whether the HGF/c-Met signaling is involved in the development of immature oocytes in PCOS. ...
... Furthermore, the comparison between the levels of HGF in FF and embryo quality indicated that the mean HGF levels in FF were significantly higher in the grade 1 embryos than those in the grades 2-4 embryos. In the ovary, HGF is known to be an autocrine/paracrine regulator of GC, theca cell growth, and steroidogenesis (10,22). Uzumcu et al. (6) demonstrated that intraovarian HGF supported folliculogenesis by mediating steroidogenesis and suppressing apoptosis in the GCs of rat ovaries. ...
Objective:
To evaluate the levels of hepatocyte growth factor (HGF) in follicular fluid (FF) and the expression of c-Met in granulosa cells (GCs) with respect to the quality of the oocyte and embryo both in patients with polycystic ovary syndrome (PCOS) and in the normal ovary during controlled ovarian hyperstimulation cycles.
Design:
Prospective controlled study.
Setting:
University hospital.
Patient(s):
Fifty-nine women undergoing IVF treatment (of whom 21 had PCOS and 38 were in the control group).
Intervention(s):
A total of 168 FF samples were collected at the time of oocyte retrieval. The HGF levels were measured by ELISA, and the mRNA expression of c-Met in GCs was detected by real-time polymerase chain reaction.
Main outcome measure(s):
The predictive values of HGF levels in serum and FF and the mRNA expression of c-Met in GCs for successful fertilization and oocyte-embryo quality.
Result(s):
The levels of HGF in serum and FF and the c-Met expression in GCs were similar between the PCOS and control groups. Granulosa cells of fertilized oocytes (2PN) had a significantly higher level of c-Met expression than that in oocytes that failed to fertilize. The mean HGF level in FF was significantly higher in the grade 1 embryos than in the grades 2-4 embryos.
Conclusion(s):
This study suggests that HGF/c-Met signaling may be a crucial determinant of fertilization success.
... Both HGF and c-met are expressed within mammalian ovaries as has been shown in the human, rat, pig, cow, and mouse ( Liu et al. 1994, Parrott et al. 1994, Parrott & Skinner 1998a, Osuga et al. 1999, Shimizu et al. 2003). Importantly, intraovarian HGF is an autocrine/paracrine regulator of GC and theca cell growth and steroidogenesis (Zachow et al. 1997, Lail-Trecker et al. 1998, Parrott & Skinner 1998b, Zachow & Woolery 2002. ...
... The steroid modulatory effects of HGF were first demonstrated using primary cultures of theca-interstitial cells from sexually immature rats. In rat theca cells in vitro, HGF suppressed LH-dependent androsterone secretion, while also blocking the steady-state level of CYP17 expression (Zachow et al. 1997). Basal and LH-induced progesterone production were stimulated in the presence of HGF; but neither the expression of CYP11A nor 3b-hydroxysteroid dehydrogenase was affected by HGF (Zachow et al. 1997). ...
... In rat theca cells in vitro, HGF suppressed LH-dependent androsterone secretion, while also blocking the steady-state level of CYP17 expression (Zachow et al. 1997). Basal and LH-induced progesterone production were stimulated in the presence of HGF; but neither the expression of CYP11A nor 3b-hydroxysteroid dehydrogenase was affected by HGF (Zachow et al. 1997). Although steroidogenesis was modulated by HGF, DNA content in rat theca cell cultures was not altered, suggesting that HGF did not stimulate proliferation of these cells. ...
The hepatocyte growth factor (HGF) system comprises HGF, its receptor (the c-met tyrosine kinase), HGF activator (HGFA) protein, and HGFA inhibitor (HAI). The components of the HGF system have been identified in a plethora of tissues to include the ovary and testis. In its traditional context, the HGF system works via paracrine- and autocrine-mediated feedback in which HGF (of mesenchymal origin) binds and activates c-met (within epithelial cells); target cells then respond to HGF via any number of morphogenic and functional changes. The concomitant presence of HGFA and HAI suggests that HGF bioactivity can be locally modulated. A number of studies have collectively shown that the mammalian ovary and testis contain HGF, c-met, and HGFA; very little is currently known regarding HAI within the gonad. Within the ovary, HGF controls numerous key functions which collectively regulate the growth and differentiation of ovarian follicles; these include cell growth, steroidogenesis, and apoptosis within theca cells and/or granulosa cells. Comparatively, less is known about the function of HGF within the testicular Leydig and Sertoli cells, but evidence is emerging that HGF may regulate somatic cell function, including Leydig cell steroidogenesis. Changes in the cellular origin of HGF and c-met during fetal and postnatal testicular development suggest that HGF, in collaboration with other growth factors, may regulate important aspects of testicular cell morphogenesis and differentiation which enable male sexual viability. Likewise, experimental evidence showing that HGF can modulate many vital processes which enable ovarian follicle growth, differentiation, and function indicate the importance of HGF in female reproduction. This review presents what is currently known regarding the expression of the HGF system and its function within the ovary and testis.
... As a result, osteoclast-specific targets to inhibit activity are lacking. Hepatocyte growth factor (HGF) is a stromal cell produced factor (Weimar et al., 1998) that causes proliferation, migration and morphogenesis of epithelial (Camussi et al., 1997; Sugawara et al., 1997; Uehara et al., 1995; Zachow et al., 1997) and endothelial (Camussi et al., 1997) cells. Moreover, the varying effects of HGF on cells are dependent on their differentiation state (Birchmeier et al., 1997). ...
Hepatocyte Growth Factor (HGF) and its protooncogene receptor c-Met regulate osteoclast function by activating pp60(c-Src) kinase and alpha(v)beta3 integrin. HGF causes transcription yet in osteoclast cells, this gene regulation is currently unknown. To begin characterization of HGF-regulated gene expression in osteoclast cells, we used a well characterized model of osteoclast cells. Using microarray, relative RT-PCR, and Western blot analyses, we have identified and confirmed differentially expressed genes in RAW 264.7 osteoclast cells in response to HGF. HGF regulation of transcription of these genes was concordant with microarray results. We report that HGF downregulates transcription factors, Distal-less 5 (Dlx-5), Distal-less 6 (Dlx-6) and Aristaless 4 (Alx-4), in RAW 264.7 osteoclast cells but has an inverse effect in undifferentiated RAW 264.7 cells.
... As a result, we used the more sensitive method of RT-PCR to analyze Mig-7 expression in 24 different human tissues (Fig. 4A). Tissues such as placenta, spleen, liver, small intestine, fetal liver, bone marrow, testis, ovary, and uterus have been shown to express both HGF and c-Met [13,14,36,41,[47][48][49][50][51][52][53][54], yet these tissues do not express detectable levels of Mig-7 transcripts even by RT-PCR. Since these pooled placenta samples in these assays were from term placentas, these results are consistent with first and second trimester placenta cytotrophoblasts cells being the most invasive in response to HGF as compared to third trimester placenta which is growth responsive to HGF [40]. ...
Hepatocyte growth factor (HGF), a cytokine involved in tumorigenesis and most metastases, initiates cell migration by binding to the protooncogene c-Met receptor. In epithelial carcinoma cells, c-Met activation causes the breakdown of E-cadherin cell–cell contacts leading to cell spreading. While the breakdown of E-cadherin contacts is immediate, HGF-induced migration requires transcription. To test the hypothesis that this de novo mRNA synthesis includes cancer cell-specific transcripts, we performed subtraction hybridization to isolate HGF-induced transcripts from an endometrial epithelial carcinoma cell line, RL95-2 (RL95), known to migrate but not to proliferate with HGF treatment. One novel cDNA we call Mig-7 is induced by HGF in endometrial epithelial carcinoma cell lines RL95 and HEC-1A before migration ensues. Ovarian, oral squamous cell, and colon metastatic tumors but not normal tissues express Mig-7. HGF did not induce Mig-7 in normal primary endometrial epithelial cells. In addition, blocking antibodies to αvβ5 integrin inhibited HGF induction of Mig-7 in RL95 cells. Most importantly, Mig-7-specific antisense oligonucleotides inhibited scattering of RL95 cells in vitro. These results are the first to demonstrate that Mig-7 expression may be used as a cancer cell-specific target to inhibit cell scattering.
... It has been generally accepted that growth of the ovarian follicle is under the endocrine control of the pituitary gonadotropins. However, a growing body of evidence indicates that steroidal and nonsteroidal factors produced by granulosa and/or theca cells affect the differentiation and proliferation of the cells located on the opposite sides of the basement membrane [1][2][3][4][5][6][7], which suggests the importance of granulosa-theca cell communication in this regulation. ...
We have investigated the role of theca cells in the control of apoptosis and proliferation of granulosa cells during bovine ovarian follicular development using a coculture system in which granulosa and theca cells were grown on opposite sides of a collagen membrane. A DNA fluorescence flow cytometry was used to determine the extent of apoptosis and proliferation in populations of granulosa cells. When granulosa cells were isolated from small follicles (3-5 mm), the percentage of apoptotic cells gradually increased by 1.8-fold during the 3 days of culture. This change was reduced (3.1-fold) by the presence of theca cells. When the cells were isolated from large follicles (15-18 mm), the percentage of apoptotic granulosa cells was gradually reduced (3.4-fold) during the 3 days of culture in single-cultured groups. The percentage of apoptosis on Day 1 was reduced (1.6-fold) by the presence of theca cells. However, such an effect was not detected on Days 2 and 3 of the culture. Theca cells did not affect the proliferation of granulosa cells obtained from either small or large follicles. The present study suggests that theca cells regulate the fate of granulosa cells throughout the follicular maturation process by secreting factors that suppress apoptosis.
... IGF-I enhances LH-stimulated CYP17 mRNA expression and androgen production in cultured rat TICs [8][9][10]24]. Stem cell factor has also been shown to increase production of androstenedione from bovine theca cells grown under confluent conditions [24]; however, transforming growth factor- [25] and hepatocyte growth factor [26] produced in vivo by ovarian thecal and interstitial cells attenuate androgen production in rat cultured thecal interstitial cells in vitro, perhaps to prevent detrimental effects on folliculogenesis [27,28]. This line of evidence suggests that the control of androgen production may include negative regulatory mechanisms, which may be important during early ovarian development. ...
Interstitial cells in the neonatal hamster do not respond to LH in vitro; however, side-chain cleavage (CYP11A1) and 17alpha-hydroxylase (CYP17) enzyme proteins are expressed in these cells. The objective of the study was to evaluate whether the cAMP second messenger system was active in these cells and if cAMP upregulates the levels of CYP11A1 and CYP17 mRNA. Interstitial cells (ICs) were cultured for 96 h in the presence of 5% fetal bovine serum and then cultured in serum-free medium in the presence of LH, forskolin, or 8-Br-cAMP for 24 h. The accumulation of cAMP, progesterone, and androstenedione was measured by radioimmunoassay, whereas CYP11A1 and CYP17 mRNA levels were determined by a semiquantitative reverse transcription-polymerase chain reaction and Southern hybridization analysis. LH failed to induce either progesterone or androstenedione production; however, forskolin stimulated cAMP production by interstitial cells in a dose-dependent manner. Moreover, both forskolin and 8-Br-cAMP significantly elevated the levels of CYP11A1 and CYP17 mRNA and induced progesterone synthesis by the interstitial cell monolayer. Despite the increase in CYP17 mRNA levels by 8-Br-cAMP, no appreciable change was noted in androstenedione production. These results suggest that, in vitro, a fully functional adenylate cyclase system is present in cultured interstitial cells of the neonatal hamster and that cAMP can influence the expression of CYP11A1 and CYP17 genes; however, cultured cells do not appear to express LH receptors that are functionally linked to the adenylate cyclase system. Moreover, the translation of CYP17 mRNA may require additional factors, which may originate from maturing granulosa cells.
