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E Fugassa

Publications and source records attributed to E Fugassa.

At least 37 records · Page 2Linked to original sources
PubMed1994

Thyroid hormone modulates the expression of rat liver gammaglutamyltranspeptidase activity.

Gammaglutamyltranspeptidase (GGT) activity is considered as a marker of liver phenotype, being maximally expressed in fetal liver and virtually absent in adult mature tissue. Since thyroid hormone has been identified to influence growth and development of almost all tissues, we have investigated the possible involvement of such factors in regulating rat hepatic GGT. Our results indicate that the activity of GGT in liver of perinatal hypothyroid rats is low as well as in control animals; instead, it is greatly increased in adult hypothyroid rats compared to controls of the same age (+260% in 2-month-old hypothyroid rats). Replacement therapy with T3 to hypothyroid rats normalizes the enzyme activity to the levels of control animals. Thyroid hormone appears to modulate the gene expression of the enzyme, since in the different thyroid status GGT mRNA level closely parallels the variations of the enzyme activity. These results further suggest that thyroid hormone plays a crucial role in maintaining the phenotype of adult liver tissue.

Animals↗
PubMed1994

Effect of thyroid status on the oxidative capacity of Sertoli cells isolated from immature rat testis.

Our previous studies indicate the Sertoli cell as a target for thyroid hormone action at testis level. In the present study we evaluated the effect of thyroid hormone on Sertoli cell oxidative capacity measured by specific cytochrome oxidase (COX) activity and intracellular adenosine triphosphate (ATP) content. Sertoli cells were isolated from 21-day-old rats. Hypothyroidism, induced from the day of birth by administration of 0.025% methimazole, was characterized by a severe delay of body and testis growth and resulted in a lower COX activity (-40%, p < or = 0.01) and a lower ATP content (-35%, p < or = 0.01) by isolated Sertoli cells. Administration of triiodothyronine (10 micrograms/100 g body wt on alternate days) to hypothyroid rats improved body and testis growth and restored both COX activity and ATP content. The presence of high-affinity, low-capacity binding sites for triiodothyronine in Sertoli cell mitochondria also was demonstrated. This study, unlike that carried out on the whole testis from adult rats, demonstrates that thyroid hormone affects the energy metabolism of Sertoli cells from midpubertal rat testes.

Adenosine Triphosphate↗
PubMed1994

Influence of thyroid hormone on androgen metabolism in peripuberal rat Sertoli cells.

The aim of the present study was to investigate the influence of thyroid hormones on androgen metabolism in Sertoli cells isolated from 3- and 4- week-old rats. Hypothyroidism was induced by the oral administration of 0.025% methimazole (MMI) from birth until the rats were killed at 3 and 4 weeks of age. Half of the MMI-treated animals were injected i.p. with L-triiodothyronine (T3 3 micrograms/100 g body weight) during the last week before death. Sertoli cells from all groups were initially cultured under basal conditions for the first 24 h and subsequently in the presence of testosterone with or without T3 for an additional 24 h. Hypothyroidism was associated with severe impairment of body as well as testicular growth. Indeed, body and testicular weights were similar in 4-week-old hypothyroid animals to those in 3-week-old control rats. Testosterone metabolism in Sertoli cells isolated from 3- and 4-week-old hypothyroid rats was mainly expressed by the lowering of 5 alpha-dihydrotestosterone + androstane 3 alpha, 17 beta-diol and an enhanced formation of 5 alpha-reduced steroids with poor androgenic properties (e.g. 5 alpha-androstane, 3, 17 alpha-dione (androstanedione), 5 alpha-androstane, 3-ol-17-one (androsterone)). Treatment of the same group of animals with T3 in vivo and in vitro did not influence the pattern of 5 alpha-reductase steroids substantially. The most striking finding in the Sertoli cells of 3-week-old hypothyroid rats was the dramatic enhancement of oestradiol formation which persisted to a lesser extent 1 week later.(ABSTRACT TRUNCATED AT 250 WORDS)

Androgens↗
PubMed1993

Thyroidal regulation of nuclear tri-iodothyronine receptors in the developing rat testis.

Previous work has demonstrated that thyroid hormones influence testis development. Specific receptors for tri-iodothyronine (T3) have been demonstrated in Sertoli cells. The aim of the present study was to examine the possible effect of thyroid hormone on its own receptor during pubertal development by evaluating the influence of thyroid status on T3-binding capacity, -binding affinity and receptor occupancy in nuclei isolated from immature rat testes. The binding capacity for T3 of nuclei from rat testis significantly decreased during pubertal development, being 375 +/- 32, 117 +/- 15 and 44 +/- 7 fmol/mg DNA in 7-, 21- and 35-day-old rats respectively, whereas the affinity of binding, as evaluated by the dissociation constant (Kd), did not change. Early induced hypothyroidism significantly affected the time-course of the postnatal decline of nuclear T3 receptors in the testis. At 21 days of age, the binding capacity for T3 in the testis of methimazole-treated rats was significantly higher with respect to euthyroid controls, being 173 +/- 21 and 117 +/- 15 mol/mg DNA respectively, while the Kd was unaffected. T3 replacement therapy completely prevented changes in T3 receptor number induced by hypothyroidism without modifying the Kd. Our results indicate that nuclear T3 receptors in the developing rat testis are modulated by thyroid hormone.

Animals↗
PubMed1993

Tri-iodothyronine effects on Sertoli cell protein turnover in the prepubertal pig.

The direct effect of Tri-iodothyronine (T3: 0.1-100 nM) on protein turnover was studied using primary cultures of Sertoli cells isolated from immature piglet testis. The results demonstrate that T3 significantly increases protein synthesis without altering the protein degradation rate. These data and previous ones, showing the presence of specific T3 receptors in Sertoli cell nuclei, indicate that T3 plays a fundamental role in the early regulation of porcine Sertoli cell growth and maturation.

Animals↗
PubMed1992

HPLC analysis of free amino acids and amino acids of total proteins in cultured cells: an application to the study of rat Sertoli cell protein metabolism.

A simple, rapid and, sensitive HPLC method, coupled with fluorometric detection, has been worked out and employed to determine the intracellular free amino acid concentrations and the amino acid composition of total proteins in rat Sertoli cell primary culture. Sertoli cells were isolated enzymatically from testes of 20- and 28-day-old rats and cultured at 32 degrees C in Eagle's minimum essential medium. On the second day of culture, cell monolayers were quickly rinsed with ice-cold saline, immediately frozen in liquid nitrogen, accurately harvested, and homogenized in 10% trichloroacetic acid. Tissue free amino acids were determined in the acidic soluble fraction following neutralization, while the precipitate was hydrolyzed for the evaluation of the fractional content of amino acids into total proteins. Amino acid samples were derivatized with o-phthaldialdehyde/3-mercaptopropionic acid and resolved by a linear one-step acetonitrile gradient in 12.5 mM sodium phosphate buffer, pH 7.2, employing a 5-microns particle size reversed-phase column. Fluorescence was monitored with excitation at 330 nm and emission at 450 nm. Under these conditions all major physiological amino acids could be satisfactory separated, identified, and subsequently quantified with the aid of standards. The run time was about 50 min; the linearity was excellent over a large range of concentrations (1-800 pmol) and the lower limit of sensitivity appeared to be 0.5 pmol. This method permits us to demonstrate age-dependent modifications in the intracellular amino acid pool and to adequately evaluate the process of protein synthesis in cultured Sertoli cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1992

Effect of epidermal growth factor on insulin-like growth factor-I (IGF-I) and IGF-binding protein synthesis by adult rat hepatocytes.

Growth hormone has been established as a primary regulator of IGF-I gene expression in adults, not only in liver but also in many extrahepatic tissues. We considered the possibility that IGF-I production by adult rat liver could also be stimulated by epidermal growth factor (EGF), a peptide known to be involved in liver regeneration. Chromatographic analysis performed after acid treatment of conditioned media revealed the presence of both immunoreactive (IR) IGF-I and IGF binding protein (IGFBP). Both IR IGF-I and IGFBP were present in the conditioned medium of adult rat hepatocytes in basal conditions. The stimulation of IGF-I and IGFBP secretion by EGF appears to be dose-dependent with a significant increment already evident at 5 nM. That EGF stimulates secretion is supported by the finding that IGF-I and IGFBP-1 mRNA levels are increased after EGF supplementation. We conclude that adult rat hepatocytes spontaneously produce IGF-I and IGFBP, and that EGF is able to increase their synthesis and secretion. This non-growth hormone-dependent regulation of IGF-I and IGFBP-1 production by adult rat hepatocytes in culture indicates an important autocrine/paracrine role for IGF-I, particularly during liver regeneration after extensive organ mass loss.

Animals↗
PubMed1992

Identification of nuclear tri-iodothyronine receptors in Sertoli cells from immature piglet testes.

The existence of nuclear tri-iodothyronine (T3) receptors in both Sertoli and Leydig cells isolated from immature piglet testes was investigated. The results demonstrated the presence of high-affinity (Kd = 1.09 +/- 0.25 nM), low-capacity (185 +/- 24 pg T3/mg DNA) binding sites for T3 in nuclei from freshly isolated Sertoli cells. No specific binding for T3 was observed in nuclei isolated from Leydig cells. The localization of specific T3 receptors, which might mediate the onset of thyroid hormone action, at Sertoli cell level confirms that these cells are a target for thyroid hormone and strongly sustain the role of the thyroid in the regulation of testicular functions during postnatal development.

Animals↗
PubMed1992

[Effect of insulin-like growth factor I (IGF-I) on Sertoli cell metabolism in the pubescent rat].

The influence of Insulin-like Growth Factor I (IGF-I) on some metabolic functions of Sertoli cells from peripubertal rats was investigated. Sertoli cells were isolated from the testes of 24-day-old animals and cultured at 32 degrees C in Eagle's MEM with or without 1 nM IGF-I. Sertoli cells cultured in the presence of IGF-I showed increased nuclear RNA polymerase activity (+80%) and augmented protein synthesis (+50%).

Animals↗
PubMed1991

Effect of thyroid hormone on the pre- and post-natal development of the rat testis.

The relationship between thyroid function and testicular development in the rat was investigated. Hypothyroidism was induced during fetal or post-natal life by adding methimazole (MMI) to the drinking water of pregnant or lactating mothers. A group of newborn rats was treated with MMI and i.p. injections of L-tri-iodothyronine (L-T3). Hypothyroidism was shown by the reduced serum levels of total T3 and of total thyroxine (T4) in pregnant mothers and in pubertal rats. Testes were studied using light microscopy at 18 and 21 days post coitum or during puberty (21, 35 and 50 days after birth); serum levels of gonadotrophins were also evaluated in pubertal rats. Hypothyroidism had no effect on testicular development during fetal life and when induced in newborn rats it was associated at puberty with reduced serum levels of FSH and LH and with delayed maturation of the testis compared with control rats. The delay in maturation consisted of a reduction in the diameter of seminiferous tubules, and a reduction in the number of germ cells per tubule; this was associated with increased degeneration and arrested maturation of germ cells. In addition, Sertoli cells demonstrated retarded development, as indicated by a delay in the appearance of cytoplasmic lipids and in the development of a tubule lumen. Hormonal and morphological abnormalities were absent in rats treated with MMI plus L-T3. In conclusion, hypothyroidism occurring soon after birth caused reduced levels of gonadotrophins in the serum and a delay in pubertal spermatogenesis, possibly due to retarded differentiation of the Sertoli cells.

Animals↗
PubMed1991

Expression of hepatic mRNAs for insulin-like growth factors-I and -II during the development of hypothyroid rats.

The effect of thyroid status on the expression of insulin-like growth factors-I and -II mRNAs in the liver of developing rats has been investigated. Northern blot analyses of the specific mRNA demonstrated the presence of four IGF-II mRNA species which were strongly expressed in fetal liver and progressively declined after birth, becoming undetectable after week 3. This decrease was markedly delayed in the liver of hypothyroid rats. In addition, expression of IGF-I mRNA, absent in fetal liver, began during week 1 after birth and progressively increased with age. This increase was markedly delayed in the liver of hypothyroid rats. The data suggest that thyroid hormones regulate rat development via the co-ordinate expression of hepatic IGF-II and IGF-I mRNAs.

Animals↗
PubMed1991

Relationship between DNA topoisomerase I activity and DNA synthesis in cultured hepatocytes: effects of orotic acid.

Previous studies have demonstrated that DNA topoisomerase I activity can be closely related to DNA replication and active transcription in different experimental models. This relationship was further investigated by studying the time course of DNA topoisomerase I activity in cultured rat hepatocytes stimulated by epidermal growth factor. This mitogen has been shown to stimulate DNA synthesis in liver cells both in vivo and in vitro. DNA topoisomerase I activity was assayed quantifying ATP-independent relaxation of a negatively supercoiled plasmid, substrate for the enzyme. Incubation of cellular extracts was carried out at 30 degrees C for 15'. Products were electrophoresed and analyzed by densitometry. DNA synthesis was measured as [3H]thymidine incorporation in the hepatocytes, after a 2 hrs pulse labelling. An increase in DNA topoisomerase I activity was observed early after epidermal growth factor addition, before the onset of DNA synthesis. Concomitant administration of orotic acid, which has recently shown mitoinhibitory effect, abolished the EGF-induced activity as well as DNA synthesis. Taken together these data indicate that, in cultured hepatocytes, the induction of DNA synthesis is supported by DNA topoisomerase I with a precise time schedule. Orotic acid administration abolishes this response and causes an overall loss in DNA topoisomerase I activity.

Animals↗
PubMed1991

[Nuclear triiodothyronine receptors in the testis of the growing rat are regulated by the thyroid status].

The effects of thyroid status on the binding capacity, association constant (Ka) and receptor occupancy during postnatal rat testis development were evaluated. Hypothyroidism (induced by oral administration of 0.05% methimazole from the day of birth) increased the total T3 binding capacity in the testis, retarding the normal developmental decrease in T3 receptor number (mean maximal binding capacities estimated by Scatchard analysis for 21-day-old eu- and hypothyroid rats were 117 and 173 fmol/mg DNA, respectively). The rat thyroid status also affected the percentage of T3 receptor occupancy but not the affinity of binding (as measured by Ka). The postnatal developmental changes in T3 binding capacity induced by hypothyroidism were completely reversed by T3 replacement. These results suggest that T3 nuclear receptors in the developing rat testis are modulated by thyroid hormones.

Animals↗
PubMed1990

Thyroid hormone stimulates the production of insulin-like growth factor I (IGF-I) by immature rat Sertoli cells.

The effects of thyroid hormone on insulin-like growth factor I (IGF-I) production by Sertoli cells isolated from immature rats have been investigated. In Sertoli cells from hypothyroid rats the production of IGF-I was significantly lower than in controls and was greatly stimulated by the administration of triiodothyronine (T3) in vivo. The in vitro addition of physiological doses of T3 (1 nmol/l) significantly increased the production of IGF-I by cultured Sertoli cells indicating a direct action of the hormone on local IGF-I production. Our results suggest the involvement of IGF-I in the thyroid hormone-dependent maturation of testicular function.

Animals↗
PubMed1990

Influence of hypothyroidism on in-vitro testicular steroidogenesis in adult rats.

The influence of hypothyroidism on testicular steroidogenesis was investigated by evaluating the production of testosterone and its precursors by isolated testes from adult male rats. Animals were made hypothyroid starting from the 4th week of life either by daily oral administration of 0.1% methimazole (MMI) or by surgical thyroidectomy (TZ). Half of the thyroidectomized rats were i.p. injected with 3 gamma T3/100 g body weight on alternate days during the last three weeks before sacrifice. Hypothyroidism is associated with a severe retardation of body growth, which appears more marked in thyroidectomized than in MMI treated rats; no significant variations in testis weight are observed. Administration of T3 does not completely restore body weight. A significant decrease in the "in vitro" production of testosterone and its precursors by testes isolated from hypothyroid rats is observed. This effect is more evident in thyroidectomized rats where a marked drop in the "in vitro" production of some testosterone delta 4 precursors is associated with the increase in DHEA/delta 4 ratio. T3 injection to thyroidectomized rats only partially restores the "in vitro" testosterone production. Results suggest that as the degree of hypothyroidism became more severe, the rate of testosterone production decreases and testicular steroidogenesis changes from the delta 4 to delta 5 metabolic pathway as a consequence of the impairement of 3-beta-ol-dehydrogenase activity.

Androstenedione↗
PubMed1990

The effect of L-acetylcarnitine on some reproductive functions in the oligoasthenospermic rat.

The effect of L-acetylcarnitine (LAC) on some parameters of male reproductive function was studied on rats made oligoasthenospermic with dibromochloropropane (DBCP). DBCP depresses sperm count and motility. After one injection of the drug, LAC induces a recovery of both sperm count and motility but after two injections it is ineffective. This effect is also shown visually by microscopic examination of seminiferous tubules. Among the enzymatic activities evaluated as biochemical markers of testicular function both lactate dehydrogenase and NADPH-cytochrome P 450-reductase increased significantly (P less than 0.05) after treatment with LAC in normal rats. LAC also stimulates testosterone production. It is suggested that LAC may affect testicular function.

Acetylcarnitine↗
PubMed1989

Thyroid hormone affects the development of Sertoli cell function in the rat.

The relationship between thyroid activity and Sertoli cell function has been investigated in prepubertal rats. Male 28-day-old Wistar rats were used to prepare Sertoli cells by sequential enzyme digestion of the testes. Hypothyroidism, induced by oral administration of methimazole from the day of birth, was characterized by a severe retardation of body and testis growth and a net inhibition of the increase in Sertoli cell gamma-glutamyl transpeptidase (GGT) activity as well as in androgen-binding protein (ABP) and lactate production, which normally occur during postnatal development of Sertoli cells. The functional parameters of Sertoli cells from hypothyroid 28-day-old rats approximated to those of cells from euthyroid 15-day-old animals. These results are consistent with the impairment of protein synthesis in Sertoli cells from hypothyroid rats compared with controls. Body and testis growth were improved and Sertoli cell functions were restored with 3,3',5-tri-iodothyronine (T3) replacement therapy. An excess of T3 in the serum, induced by daily i.p. injections of T3 (100 micrograms/kg body wt) during the last week before the rats were killed, failed to induce changes in body and testis growth or in the activity of GGT and lactate dehydrogenase of Sertoli cells. Cells from hyperthyroid rats exhibited a specific decrease in ABP production. These results indicate that thyroid hormone is necessary for the postnatal maturation of Sertoli cell function and suggest a regulatory role of the hormone on gametogenic development in the prepubertal rat.

Aging↗