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A Catizone

Publications and source records attributed to A Catizone.

17 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↗
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↗
PubMed1996

Non-specific pinocytosis by human endothelial cells cultured as multicellular aggregates: uptake of lucifer yellow and horse radish peroxidase.

We have analyzed the pattern of time-dependent and concentration-dependent incorporation of Lucifer Yellow CH (LY) and Horseradish Peroxidase (HRP) by human umbilical vein endothelial cells cultured on a non-adhesive substratum, where they they become organized into stable, multicellular aggregates. The data were compared with those previously obtained from low-density cultures of non-growing endothelial cells adherent to plastic. While the linear trend of the incorporation kinetics is preserved, the rate of uptake with both time and concentrations is highly dependent on the culture conditions, namely typology of cell-cell and cell-substrate interactions. An at least two-fold increase of the rate of uptake was observed with both markers in the aggregated cells. The extracellular concentration of LY required to saturate the binding capacity of the cell surface shifts from approximately 0.25 mg/ml, with the adherent cells, to approximately 0.5 mg/ml in the aggregated cells; the rate of uptake of three different forms of HRP shows, besides a sharp quantitative increase, also qualitative variations, testified by differential changes of their incorporation rates. These results are entirely consistent with the assumption that the association of the endothelial cells into multicellular aggregates increases the rate of pinocytic uptake by modifying the physicochemical properties of the cell surface, thereby increasing its differential affinity for the extracellular markers.

Biological Transport↗
PubMed1993

A quantitative assessment of non specific pinocytosis by human endothelial cells surviving in vitro.

Human umbilical vein endothelial cells have been assayed in vitro, 24 hrs. after plating, for non specific pinocytic activity. The culture conditions were designed to minimize the exogenous stimulations of pinocytosis, such as those possibly coming from mitotic induction and chemical and contact-dependent signaling. Two different markers were used: Lucifer Yellow CH (LY), and three different preparations of horseradish peroxidase, a multiple form (type II) composed of five different isoenzymes, and two purified acidic (type VIII) and basic (type IX) isoenzymes. The uptake of LY appears to depend on both fluid-phase incorporation and non specific adsorption to the cell surface, and it shows a linear monophasic dependence on time and a linear diphasic dependence on concentration. This probe is actively chased from the cells to an extent proportional to the amount incorporated. Therefore, the endocytic index obtained from the LY incorporation data is not a reliable estimate of extracellular fluid incorporation. The three different forms of HRP share an incorporation pattern linearly dependent on both time and concentration, consistent with the classical interpretation of a simple fluid-phase mechanism of intracellular uptake; however, the rates of uptake and chase activity of the pure isoenzymes are clearly different from that of the multiple form. The observed differences are related to possible local variations in the physicochemical properties of the cell surface, which may restrict the cell surface area suitable for fluid-phase uptake of differently charged macromolecular probes.

Cells, Cultured↗
PubMed1992

[Electric alternans of QRS: its electrophysiologic significance in narrow QRS supraventricular paroxysmal tachycardia].

The results of 22 patient's transesophageal electrophysiologic studies were analyzed to value the meaning of QRS alternans during narrow QRS tachycardia. Standard ECG showed ventricular pre-excitation in eight patients. QRS alternans were in six of them: heart rate was included between 230 and 374 m/sec (301.8 +/- 42 m/sec); in the remaining 16 patients, heart rate was included between 240 and 450 m/sec (350.0 +/- 42 m/sec). Our findings suggest that QRS alternans is related to the rate of tachycardia rather than to the presence of an accessory pathway.

Adult↗
PubMed1990

Determination of urinary tryptophan and its metabolites along the nicotinic acid pathway by high performance liquid chromatography with ultraviolet detection.

A fast and sensitive method is given for analysing urinary tryptophan and six of its metabolites on the nicotinic acid pathway. Kynurenine, tryptophan, 3-hydroxykynurenine, anthranilic acid, 3-hydroxyanthranilic acid, kynurenic acid and xanthurenic acid were isocratically eluted and completely resolved with a mobile phase of acetonitrile + sodium acetate buffer, pH 4.76 (4:96, v/v). The flow rate was 0.8 mL/min at the beginning and was then linearly increased to 1.2 after 6 min; after 14 min the flow was augmented from 1.2 to 2 mL/min. The effluent was monitored with a variable UV detector set at 254 nm for the first five peaks and at 280 and 325 nm for the penultimate peak and final peak. Analytical recoveries of the compounds after deproteinization varied between 64% and 98%. The reported method should enable one to examine easily, extensively, quantitatively and routinely urinary tryptophan and the most important metabolites of the nicotinic acid pathway.

Chromatography, High Pressure Liquid↗
PubMed1989

Epidermal growth factor binding and steroid receptor content in human benign prostatic hyperplasia.

The receptor for epidermal growth factor (EGF-R) was characterized on membrane fractions from human benign prostatic hyperplasia (BPH). Specific binding of [125I]EGF reached equilibrium after 40 min at 25 degrees C and was stable for up to 120 min. Saturation analysis of EGF-R, performed by incubating the membranes with 0.0156-15 nM [125I]EGF in the presence and in the absence of 100-fold excess of cold EGF for 60 min, revealed the presence of two classes of binding sites with high and low affinities (Kd = 0.35 +/- 0.23 and 9.60 +/- 2.87 nM respectively). Competition experiments revealed that FSH, insulin and calcitonin did not compete with [125I]EGF. The simultaneous determination of EGF-R and that of estradiol (ER), progesterone (PR) and androgen receptors (AR) was performed using the same buffer to homogenate the tissues and to obtain cellular membranes. The steroid receptors (SR) were determined by means of the dextran-coated charcoal method. There was a significant negative correlation between nuclear SR and binding capacity of EGF-R. The presence of specific and high affinity binding sites for EGF and the modulation of the level of these sites by steroid receptors suggest a possible role of EGF in prostatic hyperplasia.

Biomarkers↗
PubMedDate not supplied

Functional role of hepatocyte growth factor receptor during sperm maturation.

Mammalian spermatozoa acquire motility and fertilizing capacity during their transit through the epididymis. Hepatocyte growth factor (HGF) is a pleiotropic cytokine with potent motogenic capacities that has been identified in different organs, including the mammalian male genital tract. In mice, HGF is present in the testis and, in large amounts, in the distal part of the epididymis. In prepuberal rats, we have demonstrated that HGF is synthesized by the peritubular myoid cells and in men, HGF is present in significant quantities in seminal plasma. It has been suggested that in mice, HGF has a role in initiating sperm motility, whereas in men, no significant correlations between HGF concentration and sperm motility have been found. In the present paper we report that in rats, HGF receptor, c-met, is expressed in testicular and epididymal spermatozoa. Through immunocytochemistry, we have found that c-met is exclusively localized on the head in testicular sperm. A different localization of c-met has been found in sperm isolated from caput and cauda epididymidis. Cells isolated from epididymal caput show a c-met localization exclusively restricted to the head in most cells. In a minority of caput epididymis spermatozoa the receptor is localized both in the cell head and along the flagellum. Spermatozoa isolated from the epididymal cauda were quite homogeneous, showing the receptor localized along the entire cell surface. We also report that HGF is synthesized and secreted by the rat epididymis as indicated by the scatter effect of epididymal cell homogenate and culture medium on MDCK cells. To clarify whether HGF is involved in the acquisition of sperm motility in the epididymis, its maintenance, or both, spermatozoa isolated from caput epididymidis have been cultured in medium alone or supplemented with HGF. The results obtained indicated that HGF has a positive effect on the maintenance of sperm motility which, in the absence of HGF, significantly decreases during the first hour of culture, whereas it is maintained for at least 3 hours when HGF is present in the culture medium. We also report that HGF does not influence spermatozoa viability as indicated by the cytometrical analysis of propidium iodide-labeled sperm; an equal number of dead cells appeared in control and in HGF-treated preparations. In conclusion, our data strongly support the hypothesis that HGF positively influences sperm motility maintenance during sperm transit through the epididymis, indicating that c-met receptor and its ligand, HGF, have a role in male fertility.

Animals↗