Biomedical subjects

Vincenza Laforgia

Publications and source records attributed to Vincenza Laforgia.

At least 19 recordsLinked to original sources

PubMed2006

Molecular characterization and gene expression of the pituitary adenylate cyclase-activating polypeptide (PACAP) in the lizard brain.

The pituitary adenylate cyclase-activating polypeptide (PACAP) is considered a pleiotropic neuropeptide in vertebrate physiology. The nucleotide sequence, the expression and the distribution of PACAP were determined in the brain of the lizard Podarcis sicula. RT-PCR showed that the brain of this reptile synthesizes an mRNA coding for PACAP. By performing in situ hybridization and immunohistochemistry techniques, a wide distribution of PACAP and its mRNA in neurons, nervous fibers and other cells was found. Phylogenetic sequence analysis indicates that lizard PACAP is highly conserved, resembling the vertebrate PACAP. Our data demonstrate that PACAP is not only highly preserved during vertebrate evolution but also suggest that PACAP could be implicated in a wide number of functions in the physiology of the reptile brain.

Animals↗

PubMed2006

The adrenal gland of Triturus carnifex after glucagon administration.

The present work was undertaken in order to investigate the influence of endocrine pancreas on the adrenal gland of Triturus carnifex. Our experiments aimed at studying the effects of intraperitoneal injections of glucagon on ultrastructural morphological and morphometrical features of steroidogenic and chromaffin tissues, as well as serum levels of aldosterone, corticosterone, norepinephrine (NE) and epinephrine (E). With regard to steroidogenic tissue, in January and November, glucagon decreased lipid droplet content in steroidogenic cells, that showed clear signs of increased activity. Moreover, increased corticosteroid serum levels were found. With regard to chromaffin tissue, in January glucagon played a stimulatory role on PNMT enzyme, eliciting an increase in the presence of E granules, and a decrease in the presence of NE granules, in the chromaffin cells. Moreover, increased E serum levels and decreased NE serum levels were found. In November, glucagon gave rise to a decrease in the presence of NE and E granules in the cells; E serum levels were strongly increased, whereas NE serum levels did not undergo any significant change. These findings suggest an involvement of the endocrine pancreas of the newt in the modulation of adrenal gland activity.

Adrenal Glands↗

PubMed2006

The adrenal gland of newt Triturus carnifex (Amphibia, Urodela) following in vivo betamethasone administration.

The response of the adrenal gland of Triturus carnifex to betamethasone administration was studied; the effects were evaluated by examination of the ultrastructural morphological features of the tissues as well as the serum levels of aldosterone, corticosterone, norepinephrine and epinephrine. In March and June, betamethasone significantly decreased the serum levels of aldosterone and corticosterone and the lipid droplet content in the steroidogenic cells. Moreover, betamethasone influenced the chromaffin tissue, enhancing in March (when the chromaffin cells produce norepinephrine and epinephrine in almost equal quantities) epinephrine serum levels and the numeric ratio between norepinephrine and epinephrine granules in the chromaffin cells. In June, (when the chromaffin cells contain almost exclusively norepinephrine granules) betamethasone administration raised norepinephrine serum levels, whereas a decrease in the numeric ratio between norepinephrine and epinephrine granules in the chromaffin cells was found. Finally, betamethasone administration did not evoke in June any increase in the mean number of epinephrine granules in the chromaffin cells and/or in epinephrine serum levels, as would be expected if phenyletanolamine-N-methyl transferase (PNMT) enzyme, converting norepinephrine into epinephrine, were activated by corticosteroids. The results of this study showed that betamethasone decreased aldosterone and corticosterone serum levels and enhanced catecholamine serum concentrations. Moreover, the present results suggest that a stimulatory role of glucocorticoids on PNMT enzyme may be ruled out.

Adrenal Glands↗

PubMed2006

The newt Triturus carnifex as a model for monitoring the ecotoxic impact of the fungicide thiophanate methyl: adverse effects on the adrenal gland.

The aims of this study were to propose a bioindicator organism, the newt Triturus carnifex, for the assessment of toxicological impact of thiophanate methyl in the Campania region (Italy) and the possible adverse activity on the adrenal gland. In the acute toxicity study, experimental groups of T. carnifex were exposed to 2.40, 4.80, 9.60 and 19.20 microg/L tap water of thiophanate methyl for 2 days; the LD50 was found to be 9.60 microg/L. To evaluate the effects on the adrenal gland, newts were exposed to a dose of 25% of the LD50 2 days for 8 days. The ultrastructural features of the tissues as well as the serum levels of aldosterone, corticosterone, norepinephrine (NE) and epinephrine (E) were evaluated. The number of secretory vesicles in the chromaffin cells appeared significantly decreased, whereas NE and E serum levels appeared strongly increased. Moreover, corticosterone and aldosterone serum levels appeared significantly reduced. The results suggest that: 1) T. carnifex has the features of an ideal bioindicator, due to its high sensitivity to thiophanate methyl, 2) thiophanate methyl acts as endocrine disruptor, affecting the adrenal gland at very low doses, 3) thiophanate methyl may be toxic for nontarget organisms, such as newts.

Adrenal Glands↗

PubMed2006

Distribution of PACAP in the brain of the cartilaginous fish torpedo marmorata.

In this article, we investigated the distribution of pituitary adenylate cyclase-activating polypeptide (PACAP) and its mRNA by immunohistochemistry, in situ hybridization, and RT-PCR techniques, in the central nervous system of the elasmobranch Torpedo marmorata. RT-PCR analysis showed that the CNS of T. marmorata expresses a messenger encoding PACAP. The immunohistochemistry and in situ hybridization patterns were partly overlapping, with a major expression in the hypothalamo-pituitary region and, surprisingly, in the saccus vasculosus. Our results show that, in T. marmorata, PACAP is synthesized and widely distributed in the CNS, suggesting an as yet unidentified role for this peptide in elasmobranch brain physiology.

Animals↗

PubMed2005

Localization and role of leptin in the thyroid gland of the lizard Podarcis sicula (Reptilia, Lacertidae).

Leptin, the product of the ob gene, is a hormone secreted by adipocytes that regulates food intake and energy expenditure. The hypothalamus-pituitary-thyroid axis is markedly influenced by the metabolism status, being suppressed during food deprivation. The present study was designed to ascertain whether (1) lizard thyroid gland expresses the long form of leptin receptor (Ob-Rb) and (2) the leptin administration affects the thyroid gland activity in this species (and to verify whether leptin plays a similar role in reptiles as observed in the other vertebrates). The presence of leptin receptor in the thyroid gland of Podarcis sicula was demonstrated by immunohistochemical technique (avidin-biotin-peroxidase complex--ABC method). The role of leptin in the control of thyroid gland activity was studied in vivo using light microscopy (LM) technique coupled to a specific radioimmunoassay for thyroid-stimulating hormone (TSH) and thyroid hormones (T4 and T3). Leptin (0.1 mg/100 g body wt)/day increased T4 and T3 release for 3 days but decreased the plasma concentration of TSH; using LM clear signs of stimulation in the thyroid gland were observed. These findings suggest that systemic administration of leptin stimulates the morphophysiology of the thyroid gland in the lizard through a direct mechanism involving Ob-Rb.

Animals↗

PubMed2005

Histochemical distribution of endothelin-converting enzyme subtypes in Podarcis sicula (Squamata, Lacertidae) tissues.

The distribution of endothelin-converting enzyme (ECE) in the lizard Podarcis sicula was investigated immunohistochemically using antibodies against endothelin-converting enzyme ECE-1 and endothelin-converting enzyme ECE-2 homologues. In all the tissues examined, immunoreactivity for both antibodies was found, although the distribution and degree of expression varied. Strong immunoreactivity was found in the endothelial cells and chromaffin tissue for both enzymes, whereas other tissues such as nervous tissue, renal tissue and hepatocytes display distinct patterns. Current knowledge does not allow correlation of these distribution patterns to specific functions but the data suggest that, in reptiles as in mammals, ECE is probably involved in physiological functions such as paracrine activity through endothelins and/or other substrates.

Adrenal Glands↗

PubMed2004

Temporal expression of thyroid hormone receptor alpha1 in the liver of the lizard Podarcis sicula.

The effects of thyroid hormones on metabolism and development are mediated by thyroid hormone receptors (TRs). To gain a better understanding of the potential role of thyroid hormone receptors in the liver of the lizard Podarcis sicula, we have evaluated the expression of TRs during the more critical periods of the annual variations of thyroid activity. The results obtained have indicated that in the liver of the lizard P. sicula there are three transcripts: mRNA of 5.0 kb for TRalpha1, mRNA of 2.6 kb for TRalpha2, and 6.0 kb band, which represent unprocessed heteronuclear RNA, encoding unspliced primary transcripts of RNA prior to their processing into the mature TRalpha1 and TRalpha2. By means of slot-blot, we are able to determine that there is a change in the expression of TRs that occurs in the liver during the annual cycle of thyroid activity. A major expression registers in May, when the lizard thyroid gland shows the maximal activity. The combination of molecular biology with immunohistochemistry revealed that hepatic cells were also TRalpha IR positive. Particularly intense immunostaining was present in the cell nuclei of animals sacrified in May. These observations suggest that in lizard P. sicula the thyroid hormone (T3) might regulate hepatic activity, modulating TR mRNA levels.

Analysis of Variance↗

PubMed2004

Effects of dopamine on the adrenal gland of Podarcis sicula (Reptilia, Lacertidae).

The effects of dopamine administration on the adrenal gland of a lizard, Podarcis sicula, are described. Dopamine (0.7mg/100g body wt/day for 4 consecutive days) raised plasma ACTH and corticosterone levels (ACTH: from the basal level of 4.40+/-0.05-7.30+/-0.08pg/ml 24h after the fourth dopamine injection; corticosterone: from 3.59+/-0.03ng/ml in untreated lizards to 7.40+/-0.05ng/ml 24h after the fourth dopamine injection), showing a stimulatory effect on the pituitary-interrenal axis activity. In the chromaffin tissue dopamine apparently enhanced the activity of PNMT enzyme; in fact a strong raise in the number of adrenaline cells and a decrease in the number of noradrenaline cells were observed, decreasing the numeric NA/A cell ratio, from 1.4/1 of control specimens to 0.5/1 24h after the fourth dopamine injection. At EM level, chromaffin cells contained both NA and A granules, as well as very clear granules (CG); CG granules showed granular elements ranging between 340 and 347A in diameter. These cells might be the morphological expression of a process of catecholamine resynthesis, due to a possible increase in catecholamine release, following exposure to dopamine.

Adrenal Glands↗

PubMed2004

Distribution of apelin, the endogenous ligand of the APJ receptor, in the lizard Podarcis sicula.

Apelin is a novel bioactive peptide that has been isolated from bovine stomach extracts and identified as the endogenous ligand for the APJ receptor. Although the main physiological functions of apelin have not yet been clarified, it is known that apelin is involved in the regulation of blood pressure, central control of body fluid homeostasis and the modulation of immune response. In order to investigate the distribution of apelin in reptiles, we have performed an immunohistochemical analysis on tissues of the lizard Podarcis sicula. The peptide was found to be widely distributed, although its cellular localization differed in the various organs examined. A strong immunopositivity was found in the heart, stomach and intestine. In the spleen, an intense apelin immunopositivity was restricted to a discrete number of cells scattered throughout the red pulp and co-localized with immunoglobulin kappa and lambda chains, suggesting an analogous function of this peptide in immune responses also in reptiles. Intriguingly, apelin immunoreactivity was discretely localized in endothelial cells in the lung and thyroid gland. In the light of these data, we conclude that apelin may have multiple functions in reptiles.

Animals↗

PubMed2004

Distribution of alpha7 and alpha4 nicotinic acetylcholine receptor subunits in several tissues of Triturus carnifex (Amphibia, Urodela).

The distribution of neuronal and non-neuronal mRNAs for alpha7 and alpha4 nicotinic acetylcholine receptor subunits was investigated in Triturus carnifex tissues using the in situ hybridization approach. The findings reveal a composite pattern of expression only partially overlapping for the two subunits; subunit alpha7 seems to be expressed widely throughout nervous, gastrointestinal and skin tissues, while alpha4 is present in a restricted number of cells of nervous and gastrointestinal tissue. We also found a specific pattern for each subunit; alpha7 and alpha4 associated exclusively to the epidermal glands and hypophysis, respectively; this is probably due to alternative roles that nicotinic acetylcholine receptors play in regulating physiological functions of non-neuronal amphibian tissues, rather than as mere neurotransmitters in the nervous system.

Animals↗

PubMed2004

Neuropeptide Y modulates pituitary-adrenal axis activity in the lizard, Podarcis sicula.

The role of neuropeptide Y (NPY) in the modulation of the pituitary-adrenal axis activity in a lizard, Podarcis sicula, was investigated by in vivo NPY administration. The effects were evaluated by examination of the morphological and morphometrical features of the tissues as well as the plasma levels of ACTH, corticosterone, aldosterone, norepinephrine, and epinephrine. Intraperitoneally administered NPY (27 nmol /100g body wt) raised ACTH plasma levels (from 5.23+/-0.06 pg/ml in carrier injected specimens to 6.83+/-0.01 pg/ml, 24 h after the injection). In the steroidogenic cells a strong decrease of lipid amount was found; corticosterone plasma level increased from 6.28+/-0.02 ng/ml in carrier injected lizards to 7.96+/-0.01 ng/ml 24 h after the injection); aldosterone levels were raised from 1.88+/-0.02 ng/ml in carrier injected specimens to 6.38+/-0.05 ng/ml 24 h after the experimental treatment. In the chromaffin tissue, an increase in the number of epinephrine cells and a decrease in the number of norepinephrine cells were observed, decreasing the numeric norepinephrine/epinephrine (NE/E) cell ratio, from 1.4/1 of control specimens to 0.5/1 24 h after NPY administration. Moreover, norepinephrine plasma level were elevated from 922+/-4.30 pg/ml in carrier injected specimens to 3075+/-11.30 pg/ml 24 h after NPY administration; epinephrine plasma level increased from 502+/-2.40 pg/ml in carrier injected specimens to 2759+/-8.70 pg/ml 24 h after the experimental treatment. Consistent with these findings, morphological observations showed many chromaffin cells weakly stained and with a reduced content of secretory granules. These results suggest that, in P. sicula, NPY may play a role in the modulation of the pituitary-adrenal axis activity. Previous studies localized NPY in the epinephrine cells of P. sicula adrenal gland; taken together, these results suggest that this peptide might participate in the regulation of adrenal gland activity, enhancing corticosteroid and catecholamine secretion in a paracrine/autocrine manner. The mechanism of action of NPY is discussed.

Adrenal Glands↗

PubMed2004

Atrial natriuretic factor: localization in the adrenal gland of the lizard Podarcis sicula and effects on pituitary-adrenal axis activity.

The occurrence of atrial natriuretic factor (ANF) immunoreactivity was investigated in the adrenal gland of the lizard Podarcis sicula by avidin-biotinylated peroxidase complex (ABC) immunocytochemical technique: ANF immunoreactivity was present in the chromaffin tissue, and was absent in the steroidogenic tissue. The role of ANF in the modulation of the pituitary-adrenal axis activity was investigated in vivo by intraperitoneal administration of ANF. The effects were evaluated by examination of the morphological and morphometrical features of the tissues, as well as the plasma levels of adrenocorticotropic hormone (ACTH), corticosterone, aldosterone, norepinephrine, and epinephrine. ANF (28 microg/100 g body wt) did not affect ACTH plasma levels, that remained almost unchanged; in contrast, corticosterone plasma levels increased from 6.45 +/- 0.070 ng/ml in carrier-injected lizards to 9.69 +/- 0.080 ng/ml 24 h after the injection; aldosterone levels decreased from 2.19 +/- 0.010 ng/ml in carrier-injected specimens to 0.58 +/- 0.003 ng/ml 24 h after the experimental treatment. In the chromaffin tissue, an increase in the number of epinephrine cells and a decrease in the number of norepinephrine cells were observed, decreasing the numeric norepinephrine/epinephrine cell ratio, from 1.4/1 of control specimens to 0.3/1 24 h after ANF administration. Moreover, norepinephrine plasma levels decreased from 998 +/- 4.600 pg/ml in carrier-injected specimens to 321 +/- 2.230 pg/ml 24 h after ANF administration; epinephrine plasma levels were elevated from 614 +/- 3.410 pg/ml in carrier-injected specimens to 1672 +/- 10.800 pg/ml 24 h after the experimental treatment. The presence of ANF in the adrenal gland suggests that, also in reptiles as in other vertebrates, this peptide, locally released from the chromaffin cells, may modulate the activity of the adrenal gland, probably in a paracrine manner. The effects of ANF on the adrenal gland suggest that this peptide may affect reptilian salt and fluid homeostasis.

Adrenal Glands↗

PubMed2004

Evaluation of cyclin D1 expression and its subcellular distribution in mouse tissues.

Cyclin D1 is a key cell-cycle regulatory protein required for the cell to progress through G1 to S phase. We have shown by Western blot analysis that cyclin D1 has a wide distribution in adult mouse tissues, with its level of expression being tissue-dependent. Immunohistochemistry has also shown that cyclin D1 may be present in the cytoplasm, in the nucleus or in both these cell compartments: cytoplasmic staining was observed in both proliferating cells (e.g. kidney, intestine, stomach and salivary gland) and in the non-dividing cells (the mature neurons of adult brain), while nuclear staining was seen in the neurons of the embryonic nervous system. Immunoelectron microscopy results indicate that, in tissues where cyclin D1 is present in both compartments (e.g. intestinal enterocytes), it may move via nuclear pores from the nucleus to the cytoplasm, and vice versa. The findings as a whole suggest that cyclin D1 may play multiple roles within specific tissues, probably by interacting with different substrates, and that its transit between nuclear and cytoplasmic compartments may help maintain cell homeostasis.

Animals↗

PubMed2004

The serine protease HtrA1 is upregulated in the human placenta during pregnancy.

The placenta has a dynamic and continuous capacity for self-renewal. The molecular mechanisms responsible for controlling trophoblast proliferation are still unclear. It is generally accepted that the simultaneous activity of proteins involved in cell proliferation, apoptosis, and extracellular matrix degradation plays an important role in correct placental development. We investigated in depth the expression of the serine protease HtrA1 during pregnancy in human placenta by in situ hybridization and immunohistochemistry, we demonstrated that HtrA1 displayed a low level of expression in the first trimester of gestation and a strong increase of HtrA1 expression in the third trimester. Finally, by electron microscopy, we demonstrated that HtrA1 was localized either in the cytoplasm of placental cells, especially close to microvilli that characterized the plasma membrane of syncytiotrophoblast cells, or in the extracytoplasmic space of the stroma of placental villi, particularly in the spaces between collagen fibers and on collagen fibers themselves. The expression pattern of HtrA1 in human placentas strongly suggests a role for this protein in placental development and function. Moreover, on the basis of its subcellular distribution it can be postulated that HtrA1 acts on different targets, such as intracellular growth factors or extracellular matrix proteins, to favor the correct formation/function of the placenta.

Female↗

PubMed2003

Localization and role of serotonin in the adrenal gland of Podarcis sicula (Reptilia, Lacertidae).

The occurrence of serotonin (5-hydroxytryptamine; 5-HT) in the chromaffin cells of Podarcis sicula adrenal gland was demonstrated by immunocytochemical techniques: ABC and immunogold methods. At LM and EM levels, antiserum against 5-HT revealed serotonin immunoreactivity prevalently in noradrenalin (NA) cells, on and around secretory vesicles; adrenalin (A) cells appeared scarcely stained. The role of serotonin in the regulation of adrenal gland activity was studied in vivo using LM and EM techniques coupled to a specific radioimmunoassay for adrenocorticotropic hormone (ACTH) and corticosterone. 5-HT (0.7 mg/100 g body wt)/day for 4 days increased ACTH and corticosterone release; at LM and EM level clear signs of stimulation in the steroidogenic tissue were observed, as evidenced by the variations of lipid/cytoplasm ratio. In the chromaffin tissue, LM observations evidenced a variation of the numeric NA/A cell ratio; at EM level, chromaffin tissue showed intermediate cells with A, NA, and very clear granules with granular elements. The occurrence of these cells might be the result of a process of resynthesis following serotonin-stimulated catecholamine release. These data suggested that serotonin might be involved in the modulation of Podarcis pituitary-adrenal axis, and act as a paracrine factor to modulate corticosteroid production.

Adrenal Glands↗

PubMed2003

Response of the thyroid gland of the lizard Podarcis sicula to endothelin-1.

Although endothelins were originally discovered as peptides with vasoconstrictor activity, recent studies have indicated a number of endothelin (ET) induced hormonal functions in various tissues. We have studied the interaction of ET-1 with thyroid gland of the lizard Podarcis sicula. The effects of ET-1 administration on the plasma levels of the thyroid hormones (T3 and T4) and TSH were stimulatory. Morphological changes in the thyroid after treatment with ET-1 were also detected: the height of the epithelial cells slightly increased and the apical surface acquired microvilli protruding into the follicular lumen. The colloid filled up the lumen and showed a rich peripheral vacuolation. In conclusion, a modulatory role in the control of the reptilian thyroid gland is suggested for ET-1. This is the first report on the interaction of ET-1 with the thyroid gland of reptilian.

Animals↗

PubMed2002

Endothelin-B (ET(B)) receptor distribution in tissues of the lizard Podarcis sicula.

Although the structural and pharmacological properties of endothelin (ET) receptors have been studied, little is known concerning their physiological significance, even if each subtype is supposed to have a distinct physiological action. Thus, to further elucidate the physiological function of this receptor, we examined the presence and distribution of the endothelin-B receptor (ET(B)) subtype in tissues of the lizard Podarcis sicula, using immunoblotting and immunohistochemistry. Immunoblotting indicated that, although the ET(B) receptor appears to be ubiquitous, it is present at different levels in the tissues examined. Furthermore, immunohistochemistry showed that this receptor is very abundant in endothelial cells of all tissues, suggesting that there is an ET(B)-mediated autocrine system of endothelin, which plays an important role in the regulation of endothelial cell function. On the other hand, the presence of ET(B) immunoreactivity also in endocrine systems such as adrenal and thyroid glands suggests an involvement also in the paracrine system of these organs.

Adrenal Glands↗