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M Galdieri

Publications and source records attributed to M Galdieri.

At least 19 recordsLinked to original sources
PubMed2006

Hepatocyte growth factor modulates in vitro survival and proliferation of germ cells during postnatal testis development.

The hepatocyte growth factor (HGF) is a pleiotropic cytokine that influences mitogenesis, motility and differentiation of many different cell types by its tyrosine kinase receptor c-Met. We previously demonstrated that the c-Met/HGF system is present and functionally active during postnatal testis development. We found also that spermatozoa express c-Met and that HGF has a positive effect on the maintenance of sperm motility. In the present paper, we extend our study on the germ cells at different stages of differentiation during the postnatal development of the testis. We demonstrate that c-met is present in rat spermatogonia, pachytene spermatocytes and round spermatids and that HGF significantly increases spermatogonial proliferation in 8- to 10-day-old pre-pubertal rats. At this age HGF does not affect Sertoli cells and peritubular myoid cells proliferation. In addition, we studied the effect of the factor on germ cell apoptosis and we show that HGF prevents the germ cell apoptotic process. We also studied the effect of HGF on 18- to 20-day-old and 28- to 30-day-old rat testes. At these ages also the factor significantly increases germ cell duplication and decreases the number of apoptotic cells. However, the effect on programmed cell death is higher in the 8- to 10-day-old rats and declines in the older animals. In conclusion, we report that rat germ cells (spermatogonia, pachytene spermatocytes and round spermatids) express c-met and that HGF modulates germ cell proliferating activity and apoptosis in vitro. These data indicate that the c-Met/HGF system is involved in male germ cell homeostasis and, consequently, has a role in male fertility.

Animals↗
PubMed2006

Expression and functional role of hepatocyte growth factor and its receptor (c-met) during fetal mouse testis development.

The hepatocyte growth factor (HGF) is a pleiotropic cytokine able to regulate different cellular functions. HGF action is mediated by its receptor, c-met, a glycoprotein with tyrosine kinase activity. We previously demonstrated that c-met is expressed in the newly formed seminiferous cords of the mice embryonic testes and that HGF acts as a morphogenetic factor. In this paper, we report that at 15.5 days post-coitum (dpc) c-met is expressed in the testicular cords, whereas at 18.5 dpc c-met expression is almost exclusively localized in the interstitial tissue of the testis in particular in the fetal Leydig cells. In addition, we demonstrate that HGF gene is expressed during the fetal period of testis development, heavily detectable in the interstitial compartment of 18.5 dpc testes. Interestingly, HGF is not expressed in the Leydig cells that, as above reported, express the HGF receptor. Looking for the functional role of HGF on Leydig cells, we evaluated the amount of testosterone secreted by testes isolated from 18.5 dpc embryos and cultured in the presence of HGF. The results of the in vitro organ culture show that, at this age, HGF increases the amount of testosterone secreted in the culture medium. On the contrary, HGF does not modulate the amount of testosterone secreted by testes isolated from 15.5 dpc embryos. In conclusion, we report that HGF is produced in the interstitial compartment of the developing testis but not by the Leydig cells. Conversely, the HGF receptor c-met is expressed in the Leydig cells and HGF modulates Leydig cell function during the late period of prenatal development.

3-Hydroxysteroid Dehydrogenases↗
PubMed2005

HGF and postnatal testis development.

Hepatocyte growth factor regulates many cellular functions acting through c-met, its specific receptor with tyrosine kinase activity. We have previously reported that in prepubertal rats HGF is secreted in the seminiferous tubules by purified peritubular myoid cells whereas Sertoli cells do not express HGF mRNA. In the present paper we report that HGF is expressed by the myoid cells during the entire postnatal testicular development studied and secreted in the culture medium. On the contrary, in Sertoli cells HGF starts to be clearly detectable by northern blot at 25 days of age. HGF is expressed and secreted by Sertoli cells isolated from 35-day-old rats and is able to increase the levels of c-met expression of the Sertoli cells. Although the role of HGF during the development of the postnatal testis need further research to be clarified, the data here presented indicate that HGF is one of the growth factors regulating mammalian testicular function.

Animals↗
PubMed2004

Embryonic mouse testis development: role of platelet derived growth factor (PDGF-BB).

Platelet-derived growth factors (PDGFs) are paracrine growth factors mediating epithelial-mesenchymal interactions and exerting multiple biological activities which include cell proliferation, motility, and differentiation. As previously demonstrated, PDGFs act during embryonic development and recently, by culturing male genital ridges, we have demonstrated that PDGF-BB is able to support in vitro testicular cord formation. In the present paper, we report that PDGF-BB is present during embryonic testis development and, in organ culture, induces cord formation although with reduced diameters compared with the cords formed in the genital ridges cultured in the presence of HGF. Moreover we have analyzed the roles exerted by this growth factor during the morphogenesis of the testis. We demonstrate by immunohistochemical experiments that PDGF-BB and its receptors are synthesized by the male UGRs isolated from 11.5 and 13.5 dpc embryos and by Western blot that the factor is secreted in a biologically active form by testicular cells isolated from 13.5 dpc embryos. The biological roles of the factor have also been studied and we demonstrate that PDGF-BB acts as a migratory factor for male mesonephric cells whose migration is a male specific event necessary for a normal testicular morphogenesis. In addition we demonstrate that during testicular development, PDGF-BB induces testicular cell proliferation being in this way responsible for the increase in size of the testis. Finally we demonstrate that PDGF-BB is able to reorganize dissociated testicular cells inducing the formation of large cellular aggregates. However the structures formed in vitro under PDGF-BB stimulation never had a cord-like morphology similar to the cord-like structures formed in the presence of HGF (Ricci et al., 2002, Mech Dev 118:19-28), suggesting that this factor does not act as a morphogenetic factor during testicular development. All together the data presented in this paper demonstrate that PDGF-BB and its receptors (alpha- and beta-subunits) are present during the crucial ages of embryonic mouse testis morphogenesis and indicate the multiple roles exerted by this factor during the development of the male gonad.

Animals↗
PubMed2004

Microgravity effect on testicular functions.

In mammals spaceflight influences spermatogenesis since spermatogonial germ cell proliferation, compared to synchronous controls, is lightly decreased in irradiated or flown rats. Moreover, changes of the plasmatic testosterone production was described either in flight rats, or in rats maintained in simulated microgravity conditions. The hormonal levels of the astronauts change as it has been previously described, including hormones involved in the regulation of spermatogenesis such as testosterone and luteinizing hormone (LH). In microgravity conditions, human testosterone levels decreased whereas circulating LH levels increased. To study the effect of simulated microgravity on mammalian spermatogenesis we have utilized the Rotary Cell Culture System (RCCS) and we have cultured testicular fragments isolated from prepuberal rats in a chemically defined medium for three days under microgravity conditions. As control we have cultured the same amount of fragments at unit gravity. The morphology of the samples has been studied and the number of proliferating cells has been counted in control samples and in samples maintained in RCCS. The results indicate that the number of duplicating cells in the tubules was significantly increased in the microgravity-cultured fragments. The amount of testosterone secreted in the culture medium has been also evaluated and in RCCS samples the amount of the hormone was higher respect to the control samples.

Animals↗
PubMed2002

Pleiotropic activity of hepatocyte growth factor during embryonic mouse testis development.

The hepatocyte growth factor (HGF) is a pleiotropic cytokine whose action is mediated by c-met, a glycoproteic receptor with tyrosine kinase activity which transduces its multiple biological activities including cell proliferation, motility and differentiation. During embryonic development HGF acts as a morphogenetic factor as previously demonstrated for metanephric and lung development. Recently, culturing male genital ridges, we demonstrated that HGF is able to support in vitro testicular cord formation. In the present paper we report the expression pattern of the HGF gene during embryonic testis development and the multiple roles exerted by this factor during the morphogenesis of this organ. Northern blot analysis reveals a positive signal in urogenital ridges isolated from 11.5 days post coitum (dpc) embryos and in testes isolated from 13.5 and 15.5 dpc male embryos. On the contrary HGF mRNA is undetectable in ovaries isolated from 13.5 and 15.5 dpc embryos. Moreover, we demonstrate that HGF is synthesized and secreted by the male gonad and is biologically active. These data indicate a male specific biological function of HGF during embryonic gonadal development. This hypothesis is supported by the in vitro demonstration that HGF acts as a migratory factor for male mesonephric cells which is a male specific event. In addition we demonstrate that during testicular development, HGF acts as a morphogenetic factor able to reorganize dissociated testicular cells which, under HGF stimulation, form a tridimensional network of cord-like structures. Finally, we demonstrate that HGF induces testicular cell proliferation in this way being responsible for the size increase of the testis. All together the data presented in this paper demonstrate that HGF is expressed during the embryonic development of the testis and clarify the multiple roles exerted by this factor during the morphogenesis of the male gonad.

Animals↗
PubMed2001

Expression and functional role of hepatocyte growth factor receptor (C-MET) during postnatal rat testis development.

The met protooncogene encodes the hepatocyte growth factor receptor (HGFR, c-met). C-met, a tyrosine kinase receptor protein, is widely expressed in different cell types including the male reproductive tract. As we recently demonstrated, both c-met messenger RNA and protein are expressed in prebuberal rat testis. The aim of this work was to detect the expression of c-met during postnatal testis development and to study its functional role. Our findings show that in total rat testis c-met is expressed during postnatal life until the sexual maturation of the animals. To evaluate the receptor expression in the different cell types in the testis, homogeneous cell populations of Sertoli and peritubular myoid cells were isolated from the seminiferous tubules of 10- and 35-day-old animals. c-met gene is expressed in myoid cells at the ages considered and its expression decreases with increasing age. By contrast, in Sertoli cells c-met expression is first detectable at 25 days of life and its expression increases with the increasing age being well evident at 35 days of age. C-met protein was detected by immunocytochemistry and its expression correlates with gene expression. The receptor is functionally active because HGF administration induces morphological changes in myoid cells and in c-met-expressing Sertoli cells. As a consequence of HGF addition, Sertoli cells cultured on reconstituted basement membrane reorganize into cord-like structures that resemble testicular seminiferous cords. The data here reported demonstrate for the first time that in Sertoli cells c-met expression is developmentally regulated being present and functionally active in postpuberal Sertoli cells. Given that c-met expression persists in myoid cells during postnatal testis development and that in Sertoli cells its expression correlates over time with germ cell differentiation and lumen formation, we conclude that the c-met/HGF system is involved in testis development and function.

Animals↗
PubMed2000

Retinoid modulation of plasminogen activator production in rat Sertoli cells.

Tissue type (t) and urokinase type (u) plasminogen activators (PAs) have been shown to be secreted by Sertoli cells in the seminiferous tubules in a cyclic fashion and to be dependent upon FSH stimulation or upon the presence of adjacent spermatogenic cells. In the present study we have analyzed the production of PAs by retinoid-treated rat Sertoli cells. In addition, because retinoids modulate the response of Sertoli cells to FSH either potentiating or antagonizing its action, we have investigated a possible modulation of FSH-stimulated PA production. Under basal conditions, Sertoli cells, isolated from prepubertal rats, secrete predominantly uPA. A significant dose-dependent inhibition of uPA activity was observed after treatment with retinol, while no significant effect was detected upon tPA secretion. When Sertoli cells were cultured in the presence of 0.25 microM retinol, a significant inhibition of uPA activity was evident after 16 h of treatment and reached approximately 80% after 48 h of treatment. The analysis of the mRNA levels revealed that retinol induces an inhibition of the steady-state levels of uPA mRNA without affecting those of tPA. Moreover, retinol affected uPA mRNA levels by increasing mRNA turnover. The effect of retinoids on Sertoli cells isolated from older animals was less evident, possibly due to the reduced production of uPA with the increase of age of the donor animals. Our results on the effect of retinoids upon Sertoli cell uPA production reinforce the importance of retinoids in the control of postnatal testis development.

Aging↗
PubMed2000

Prevention of spontaneous autoimmune diabetes in NOD mice by transferring in vitro antigen-pulsed syngeneic dendritic cells.

To evaluate the effect of antigen-pulsed dendritic cell (DC) transfer on the development of diabetes, 5-week-old female NOD mice received a single iv injection of splenic syngeneic DC from euglycemic NOD mice pulsed in vitro with human y globulin (HGG). Eleven of 12 mice were protected from the development of diabetes up to the age of 25 weeks, and the insulitis score was significantly reduced. In contrast, NOD mice receiving unpulsed splenic DCs showed histological signs of insulitis and course of type 1 diabetes similar to untreated NOD mice. Treatment with HGG-pulsed DC was associated with profound modifications of cytokine secretory capacities within the islets. Thus, supernatants of islets from these mice contained increased levels of interleukin (IL)-4, IL-10, and, to a lesser extent, interferon-gamma and diminished levels of tumor necrosis factor-a compared with controls. Because exogenous IL-4 and IL-10 exert antidiabetogenic effect in NOD mice and early blockade of endogenous tumor necrosis factor-alpha prevents NOD mouse diabetes, these phenomena may be causally related to the antidiabetogenic effect of HGG-pulsed DC treatment.

Animals↗
PubMed2000

Multiple low-dose and single high-dose treatments with streptozotocin do not generate nitric oxide.

Streptozotocin (STZ) is a widely used diabetogenic agent that damages pancreatic islet beta cells by activating immune mechanisms, when given in multiple low doses, and by alkylating DNA, when given at a single high dose. Actually, STZ contains a nitroso moiety. Incubation of rat islets with this compound has been found to generate nitrite; moreover, photoinduced NO production from STZ has been demonstrated. These reports have suggested that direct NO generation may be a mechanism for STZ toxicity in diabetogenesis. Several other studies have denied such a mechanism of action. This study has shown that (1) the multiple low-dose (MLDS) treatment does not stimulate NO production at the islet level; in fact, nitrite + nitrate levels and aconitase activity (also in the presence of an NO-synthase inhibitor, namely NAME) remain unmodified; RT-PCR analysis demonstrates that this treatment does not stimulate iNOS activity; (2) the high-dose (HDS) treatment does not stimulate NO production; in fact nitrite + nitrate levels remain unmodified and iNOS mRNA levels are not altered, although aconitase activity is significantly decreased. Moreover, we have confirmed that the MLDS treatment is able to decrease SOD activity by day 11 and that STZ, given in a single high dose, transiently increases superoxide dismutase (SOD) values (24 h from the administration), then dramatically lowers SOD levels. On the basis of our results, we conclude that STZ, "in vivo" is unable to generate NO, both as a MLDS or HDS treatment, thus excluding that NO exerts a role in streptozotocin-dependent diabetes mellitus.

Animals↗
PubMed1999

Hepatocyte growth factor (HGF) receptor expression and role of HGF during embryonic mouse testis development.

The hepatocyte growth factor (HGF) receptor, c-met, transduces the HGF multiple biological activities. During embryonic development the system HGF/c-met regulates the morphogenesis of different organs and tissues. In this study we examined c-met gene expression during mouse testis development and, by means of Northern blot and in situ hybridization, we report the receptor expression pattern. C-met expression is not detectable in male genital ridges isolated from embryos at 11.5 days postcoitum (dpc). In testes isolated from 12.5 and 13.5 dpc, c-met expression is detectable and essentially localized in the developing cords. Male genital ducts do not express c-met at the reported ages, whereas female ducts appear c-met positive. Moreover, we report that HGF is able to induce testicular morphogenesis in vitro. Male genital ridges isolated from embryos at 11.5 dpc are morphologically nonorganized. Culturing 11.5 dpc urogenital ridges in the presence of HGF we obtained testis organization and testicular cord formation. Our data demonstrate that c-met is expressed during the beginning period of testis differentiation and that HGF is able to support testicular differentiation in vitro. All these data indicate that this growth factor, besides its role as mitogenic factor, plays a fundamental role during testicular cord formation probably inducing cell migration and/or cell differentiation.

Animals↗
PubMed1999

Vitamin A modulation of basement membrane production by purified testicular myoid cells.

Purified myoid cells, isolated from prepubertal rat testes, cultured in a chemically defined medium for up to 1 week do not change their metabolic activities, evaluated as protein synthesis and secretion, during the culture time. We report that fibronectin, collagen IV, and laminin are synthesized and secreted by myoid cells as demonstrated by immunocytochemical and biochemical methods. The deposition of all three proteins was spotty, with different regional localizations. The effect of vitamin A on the synthesis and the secretion of the basement membrane components was also evaluated. Retinol supplementation induces a higher synthesis of fibronectin and laminin, whereas it does not change collagen IV synthesis and secretion. The secretion of the other two molecules is differentially regulated by retinol; in fact fibronectin secretion is increased, whereas laminin secretion is reduced. Similar results were obtained utilizing retinoic acid. The data we report in this paper show, for the first time, that purified testicular myoid cells synthesize and secrete fibronectin, collagen IV, and laminin and that synthesis and secretion of these components of the basement membrane are regulated by retinol. These findings reveal a new effect of vitamin A in the regulation of mammalian spermatogenesis.

Animals↗
PubMed1999

Hepatocyte growth factor and c-MET are expressed in rat prepuberal testis.

The hepatocyte growth factor (HGF) receptor (c-MET) is present in different mammalian tissues and transduces multiple biological effects. The HGF is known to regulate many fundamental cellular functions, such as cell growth, movement and differentiation, and is involved in embryonal morphogenesis. We have studied HGF and c-MET expression in prepuberal rat testis. c-MET gene expression was found in total testis and in homogeneous cell populations, as demonstrated by Northern blotting. In the seminiferous tubules, c-MET gene was only expressed in the myoid cells. In these cells, c-MET was detectable and constantly expressed for at least six days of culture. The interstitial tissue was also c-MET positive. The protein encoded by the MET proto-oncogene was detected in myoid cells, and HGF administration to these cells induced morphological changes in the cells. HGF expression was not detected by Northern blotting using RNA extracted from total testis. By contrast, when homogenous cell populations were used, HGF expression was detectable and exclusively localized in myoid cells. Myoid cell-conditioned medium was able to induce scattering of canine kidney epithelial (MDCK) cells, and the scatter effect of a 3-days conditioned medium was evident even after 7-fold dilution of the medium. Our findings demonstrate that HGF and its receptor are present in rat prepuberal testis. The coexpression of factor and receptor in the myoid cells suggests a new role for HGF as autocrine regulator of myoid cell function and, possibly, as regulator of mammalian testicular function.

Animals↗
PubMed1998

Characterization of different cell populations isolated from rat testis peritubular cells.

Peritubular cells, a mixed population of myoid and non-myoid cells obtained by enzymatic treatments of rat seminiferous tubules, are currently utilized as a myoid cell population. We have morphologically and biochemically compared peritubular cells and purified myoid cells isolated from prepubertal rats. The two cell populations appear morphologically similar when cultured in the presence of serum: both appear to be composed of elongated cells when observed by phase-contrast microscopy. However, the two populations differ in the percentage of alkaline phosphatase positive cell since only a minority of the cells present in the peritubular cell population are positive for alkaline phosphatase expression. Secretory activity of myoid cells is higher when compared to the peritubular cell secretion. To the contrary, fibronectin is highly synthesized and secreted by the peritubular cells, suggesting that the non-myoid cells synthesize fibronectin at a high level. Myoid cells have also been cultured in a chemically defined medium without any serum addition. In this experimental condition the cells show a polygonal shape, which remains constant during culture time. Secretory activity and fibronectin synthesis and secretion of the cells cultured without serum are lower when compared to the values obtained in the presence of serum at the beginning of the culture. Moreover, the distribution of the cell-associated fibronectin is modified by the culture conditions: spotted on cells cultured in the absence of serum and fibrillar on cells cultured with serum. We therefore conclude that peritubular cells and myoid cells are significantly different cell populations and that serum addition to the culture medium dramatically influences the morphology and the metabolic activities of the myoid cells.

Animals↗
PubMed1996

Retinoic acid synthesis in the somatic cells of rat seminiferous tubules.

At physiological plasma concentrations, retinoic acid (RA) cannot cross the blood-testis barrier formed by Sertoli and peritubular cells, and it is thought to be mainly synthesized in situ through the oxidation of retinol. We have thus examined the in vitro RA biosynthetic capacity of cultured Sertoli and peritubular cells isolated from the seminiferous tubules of prepubertal rats, using holo-cellular retinol binding protein (CRBP) as a substrate. Although both somatic cell types contain CRBP and retinoic acid nuclear receptors, RA synthesis was only detected with Sertoli cell subcellular fractions. Most of the RA synthesizing activity of these cells is contributed by a microsomal-cytosolic system that shares many functional similarities with a RA biosynthetic pathway originally identified in rat liver. RA synthesis is maximal at a time of postnatal life (20 days) preceding meiotic cell accumulation and remains nearly constant thereafter. The unique ability of Sertoli cell subcellular fractions to support RA formation from holoCRBP, along with the observed age-dependent modulation of this activity, indicate that Sertoli cells represent the main site of intratubular RA production and that they may play a key role in controlling RA-dependent processes within the seminiferous tubule.

Age Factors↗
PubMed1996

Rat peritubular cell secretory activity: modulation by vitamin A.

Protein synthesis and secretion by rat peritubular cells cultured in the presence or in the absence of vitamin A were analyzed after labelling the cells with different amino acids. Vitamin A reduces the amount of proline incorporated into the newly-synthesized proteins secreted by the peritubular cells. By mono- and two-dimensional gel electrophoresis analysis, qualitative differences induced by retinol addition were detected in the proteins secreted by the proline-labelled cells. In particular we demonstrate that the amount of fibronectin secreted by the cells is reduced after retinol addition. Northern blot analysis indicates that fibronectin messenger RNA transcription is also reduced as a consequence of the vitamin addition.

Animals↗
PubMed1994

Retinoids regulate gonadotropin action in cultured rat Sertoli cells.

The effect of retinoids on cultured rat Sertoli cells was studied by evaluation of cAMP and estradiol production after gonadotropin stimulation in the presence or absence of the retinoid. Sertoli cells cultured in the presence of FSH produce a high amount of cAMP and increase their aromatase activity. The addition of retinol alone has no effect on cAMP and estradiol production; however, the presence of retinol in the culture medium exerts an inhibitory effect on Sertoli cell response to FSH stimulation. In particular, FSH-induced cAMP production of rat Sertoli cells was significantly reduced (50-60% decrease) both by retinol and by retinoic acid. This effect was observable during the first ten days of culture and was also evident when Sertoli cells were cultured in the presence of retinol and methylisobutylxanthine, an inhibitor of phosphodiesterase activity. Cholera toxin-stimulated cAMP levels were reduced by retinol, whereas forskolin-induced elevation of cAMP levels was not affected by vitamin treatment. The inhibitory effect of retinoids on FSH-stimulated aromatase activity of Sertoli cells, which is cAMP mediated, was also evident. In conclusion, the present study demonstrates that retinoids modulate FSH action on cultured rat Sertoli cells and decrease cAMP production.

1-Methyl-3-isobutylxanthine↗