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S Ceccatelli

Publications and source records attributed to S Ceccatelli.

At least 55 records · Page 3Linked to original sources

Expression of enkephalin and dynorphin precursor mRNAs in brain areas of hypo-and hyperthyroid rat: effect of kainic acid injection.

An abnormal thyroid status induces morphological and neurochemical modifications in the adult brain. In this study we have analyzed the expression of enkephalin (ENK) and dynorphin (DYN) precursor mRNAs by means of in situ hybridization in the brain of hypothyroid and hyperthyroid rats. The influence of thyroid hormones on kainic acid (KA)-induced expression of ENK and DYN mRNAs in the granule cells of the dentate gyrus was also studied. Our results can be summarized as follows: (1) hypothyroidism induces an up-regulation of ENK mRNA in the granule cells of the dentate gyrus and layers V/VI of the cingulate cortex and of DYN mRNA in the granule cells of the dentate gyrus; (2) the up-regulation of ENK mRNA expression in the granule cells induced by KA is not modified by altered thyroid status; (3) in contrast, the KA injection fails to up-regulate DYN precursor mRNA expression in the granule cells of the rostral dentate gyrus of the hypothyroid rats; (4) injection of KA in hyperthyroid rats increases the expression of DYN mRNA in the granule dentate gyrus more than in euthyroid rats. The present results suggest that thyroid hormones exert an inhibitory control of expression of ENK and DYN mRNAs in selected brain areas. This effect could be directly mediated though the thyroid hormone nuclear receptor or could be secondary to changes in glutamatergic transmission in the dentate gyrus, as suggested by the profound alteration of the KA-induced expression of DYN mRNA in the dentate gyrus of hypo-and hyperthyroid rats.

Animals↗

Adrenocortical apoptosis in hypophysectomized rats is selectively reduced by ACTH.

Using in situ end-labelling of DNA breaks (TUNEL) and staining with propidium iodide, we have investigated the occurrence of apoptosis in the adrenal cortex of hypophysectomized rats. Apoptosis was detected in the zona fasciculata and reticularis 3 days after hypophysectomy. Six days after hypophysectomy rats showed apoptotic cells mainly in the zona fasciculata. With the exception of the zona reticularis, cell death was largely prevented by ACTH replacement in 3-day hypophysectomized animals. The present results demonstrate that ACTH can reduce but not fully prevent apoptosis in the adrenal cortex of hypophysectomized rats and suggest that lack of other factors, such as other pituitary hormones, may be involved in the onset of apoptosis in the zona reticularis.

Adrenal Cortex↗

Colchicine induces apoptosis in cerebellar granule cells.

Exposure to 1 microM colchicine, a microtubule disrupting agent, triggered apoptosis in rat cerebellar granule cells (CGC). Apoptotic nuclei began to appear after 12 h followed by oligonucleosomal DNA laddering, whereas inhibition of the mitochondrial 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide metabolism became significant between 18 and 24 h, when most cells already had apoptotic nuclei. These events were preceded by loss of tau protein and fragmentation of alpha and beta tubulins. Colchicine treatment also caused alterations in Ca2+ responses to chemical depolarization and a moderate, but progressive, increase in the resting intracellular Ca2+ concentration. Nearly all neurons expressed c-Fos after the treatment with colchicine. However, while in part of the cell population c-Fos levels subsequently declined, in the neurons undergoing apoptosis the protein was still expressed, but had an abnormal intracellular localization. An increased expression of the constitutive nitric oxide synthase (NOS-I) was also detected at 12 h and was followed by increased nitrite production. Treatment with 100 nM taxol to stabilize the microtubuli prevented DNA laddering and apoptotic body formation induced by colchicine. In contrast, pretreatment with the N-methyl-D-aspartate receptor-antagonist, MK-801, or L-type Ca2+ channel blockers did not prevent colchicine-induced CGC apoptosis. Inhibitors of NOS were also ineffective in preventing apoptotic body formation and DNA laddering, whereas they delayed the secondary cell lysis. These results support the idea that colchicine-induced cytoskeletal alterations directly initiate the genetic and structural modifications that result in CGC apoptosis.

Amino Acid Oxidoreductases↗

Immunohistochemical demonstration of nitric oxide synthase in the peripheral autonomic nervous system.

In the present immunohistochemical study the distribution of nitric oxide synthase (NOS) was studied in various autonomic ganglia and in related peripheral tissues of the rat. For comparison some other neuronal markers including acetylcholinesterase and tyrosine hydroxylase as well as several neuropeptides were analysed on adjacent or the same sections. The distribution of NOS-like immunoreactivity (LI) and of these other markers has been semiquantitatively summarized. In some parasympathetic ganglia such as the sphenopalatine and submandibular ganglia NOS-LI was present in most ganglion cells, at least partly coexisting with peptide histidine isoleucine (PHI), vasoactive intestinal polypeptide (VIP) and neuropeptide tyrosine (NPY). In the pelvic ganglia a comparatively smaller proportion of neurons was NOS-positive and they often contained VIP-LI and less frequently NPY-LI. In the tissues innervated by these ganglia, such as nasal mucosa and salivary glands, NOS-positive fibers were observed around blood vessels and within the glandular parenchyma, although generally less abundant than VIP/PHI nerves, while in the uterus, vas deferens and penis a more close correlation was seen. NOS-positive fibers were also widely distributed in other tissues. In the sympathetic ganglia NOS-LI was mainly present in dense fiber networks, which disappeared after transection of the sympathetic trunc central to the ganglion. Since many cell bodies in the sympathetic lateral column of the spinal cord also were NOS-positive, it is likely that the majority of preganglionic fibers innervating sympathetic ganglia are NOS-positive. VIP-positive cells in stellate ganglia did not contain NOS-LI. The present results suggest that NO may be a messenger molecule both in parasympathetic postganglionic neurons and in preganglionic sympathetic neurons.

Amino Acid Oxidoreductases↗

Upregulation of nitric oxide synthase and galanin message-associated peptide in hypothalamic magnocellular neurons after hypophysectomy. Immunohistochemical and in situ hybridization studies.

The expression of several bioactive molecules in magnocellular hypothalamic neurons is modified when the axons of these cells are transected. In this study we have evaluated by means of immunocytochemistry and in situ hybridization the effect of hypophysectomy on the expression of nitric oxide synthase (NOS)- and of galanin message-associated peptide (GMAP)-like immunoreactivities (-LIs) as well as on their respective mRNAs in hypothalamic magnocellular neurosecretory neurons. The results show a transient increase in NOS- and GMAP-LIs in magnocellular neurons of both the paraventricular and supraoptic nuclei when compared to normal animals. The maximal increase in staining was observed between 5 and 7 days, and by 14 days NOS-LI was back to normal levels, whereas strong GMAP-LI could still be detected in a few cells. A similar picture was observed for the NOS and GMAP mRNAs. The functional significance of the present findings is unclear, but they indicate a possible role of nitric oxide and GMAP in neurosecretory neurons after injury.

Amino Acid Oxidoreductases↗

Nitric oxide synthase increases in hypothalamic magnocellular neurons after salt loading in the rat. An immunohistochemical and in situ hybridization study.

Magnocellular hypothalamic neurons of the paraventricular (PVN) and supraoptic (SON) nuclei have been shown to contain a wide variety of messenger molecules in addition to vasopressin and oxytocin, including the nitric oxide (NO)-synthesizing enzyme (NOS). In this paper we have investigated the effects of salt loading on the expression of NOS by means of immunohistochemistry and in-situ hybridization. The results show an increase in the number of NOS-immunoreactive (IR) neurons both in the PVN and the SON after 5 and 14 days of salt loading. Several of these neurons were double labelled with vasopressin antiserum. In situ hybridization showed a marked increase in the number of neurons expressing NOS mRNA and a stronger signal in individual neurons. The present results suggest a role for NO in the magnocellular hypothalamic system after salt loading.

Amino Acid Oxidoreductases↗

Plasticity of NO synthase expression in the nervous and endocrine systems.

Using immunohistochemistry and in situ hybridization the effect of nerve injury and of hormones was analysed in sensory and hypothalamic systems and in the pituitary gland. After peripheral axotomy a marked increase in NOS protein and mRNA levels was observed in dorsal root ganglia, the trigeminal ganglion and a less dramatic effect in the nodose ganglia. This effect lasted in the dorsal root ganglion neurons for at least 10 weeks. In the hypothalamic magnocellular neurons a transient increase was observed in the paraventricular and supraoptic nuclei. A similar effect was also seen after salt loading. In the anterior pituitary gland NOS was expressed in gonadotrophs and folliculo-stellate cells. Castration markedly increased NOS levels in the anterior lobe, and this could be counteracted by steroid hormone replacement. Thus, the present results show that the constitutive, neuronal NOS can be dramatically regulated in response to various manipulations, suggesting an important involvement of NO in these situations.

Amino Acid Oxidoreductases↗

Regulation of VIP mRNA expression by thyroid hormone in different brain areas of adult rat.

A role of thyroid hormone in the regulation of neuropeptide synthesis has been demonstrated in different tissues. In this paper we investigated the vasoactive intestinal peptide (VIP) mRNA expression by means of in situ hybridization in several brain areas of hypo- and hyperthyroid adult rats. Neither hypo- nor hyperthyroidism modified the VIP mRNA levels in the thalamus and in the hypothalamic suprachiasmatic nucleus. In contrast, in the anterior cingulate and frontoparietal motor cortex of hypothyroid rats there was a marked increase in the signal for VIP mRNA per cell, but the number of VIP expressing neurons did not change. These data indicate that also central VIP synthesis can be influenced by the levels of circulating thyroid hormone, but that this effect is confined to specific areas and cell populations of the brain.

Animals↗

Nitric oxide synthase in the rat anterior pituitary gland and the role of nitric oxide in regulation of luteinizing hormone secretion.

By using immunohistochemistry and in situ hybridization, we have demonstrated that the nitric oxide (NO)-synthesizing enzyme NO synthase is present in gonadotrophs and in folliculo-stellate cells of the anterior pituitary gland of male and female rats. A marked increase in levels of NO synthase protein and mRNA was observed after gonadectomy. In vitro studies on dispersed anterior pituitary cells suggest that NO inhibits gonadotropin-releasing-hormone-stimulated luteinizing hormone release. An inhibitory effect of NO has also been shown on growth-hormone-releasing-hormone-stimulated release of growth hormone [Kato, M. (1992) Endocrinology 131, 2133-2138]. Thus these findings support a dual mechanism for NO in the control of anterior pituitary hormone secretion, an autocrine mediation of luteinizing hormone release on gonadotrophs, and a paracrine effect on growth hormone secretion involving folliculo-stellate cells closely related to somatotrophs. We speculate that NO may participate in producing the pulsatile secretion patterns of these two pituitary hormones.

Amino Acid Oxidoreductases↗

The effect of lactation on nitric oxide synthase gene expression.

Nitric oxide-synthesizing (NOS) enzyme has been identified in several neural populations, including the hypothalamic paraventricular nucleus (PVN). The PVN plays a major role in regulating milk ejection. In the present study, using in situ hybridization, the effect of lactation on NOS mRNA in the PVN was investigated. A significant increase was seen in the PVN of lactating rats, indicating a possible involvement of nitric oxide in the hypothalamic-pituitary regulation of milk ejection.

Amino Acid Oxidoreductases↗

CGRP-like immunoreactivity in A11 dopamine neurons projecting to the spinal cord and a note on CGRP-CCK cross-reactivity.

Using the indirect immunofluorescence technique and double labelling procedures combined with retrograde tracing it could be demonstrated that the A11 dopamine cell group, located at the border between the diencephalon and mesencephalon of the rat brain and some of which project to the spinal cord, contains calcitonin gene-related peptide (CGRP)-like immunoreactivity. Thus, another catecholamine group in the rat brain has been shown to have a coexisting peptide. One of the CGRP antisera used in the present study also stained cholecystokinin (CCK) containing neurons in various brain areas. Absorption and displacement experiments using immunohistochemistry and radioimmunoassay showed that this cross-reactivity was confined to the C-terminal portion of the peptide molecule. Therefore, the present results suggest that CGRP antisera used for immunohistochemistry and radioimmunoassay should be tested for possible cross-reactivity with CCK.

Animals↗

NOS mRNA in the paraventricular nucleus of young and old rats after immobilization stress.

Nitric oxide synthesizing (NOS) enzyme has been localized in distinct neural populations, including the hypothalamic paraventricular nucleus (PVN). Using in situ hybridization, we have investigated possible changes in NOS mRNA in the PVN of young and old rats after 2 h of immobilization stress. In old rats a 3-fold increase in NOS mRNA was observed in the PVN immediately after the stressor, whereas in young rats the increase reached significance when animals were killed 3 h after the end of the stress procedure. The present results suggest that nitric oxide in the PVN may be involved in the stress response. The differences observed in NOS mRNA between young and old rats could be related to an age-dependent different responsiveness to stressful stimuli.

Aging↗

Effect of different types of stressors on peptide messenger ribonucleic acids in the hypothalamic paraventricular nucleus.

Using in situ hybridization we have studied the effects of different types of stressors, such as ether, immobilization, cold and swimming, on the expression of several peptide messenger ribonucleic acids (mRNAs) in the hypothalamic paraventricular nucleus of adult male rats. Paraventricular nucleus sections were hybridized using synthetic oligonucleotide probes complementary to mRNA for corticotropin-releasing hormone, neurotensin, enkephalin and thyrotropin-releasing hormone. A clear upregulation of neurotensin mRNA was seen after ether and, to a lesser extent, after immobilization stress, whereas after the two other stressors neurotensin mRNA was undetectable, as in control rats. An increase in enkephalin mRNA was observed in a selective region of the dorsal part of the medioparvocellular subdivision of the paraventricular nucleus only after ether and immobilization stress. No significant changes were seen in corticotropin-releasing hormone and thyrotropin-releasing hormone mRNA levels in any of the experimental paradigms. The present results show selective changes for various peptide mRNAs in the paraventricular nucleus after various types of stress. Significant effects could be demonstrated only on neurotensin and enkephalin mRNA after ether and immobilization stress. This suggests that adaptive changes in the rate of synthesis, processing and transport of the peptide may develop over a longer period of time.

Animals↗

Presence of a dynorphin-like peptide in a restricted subpopulation of catecholaminergic neurons in rat nucleus tractus solitarii.

Immunofluorescence colocalization techniques were used to examine the extent of coexistence of the endogenous opioid peptide dynorphin with catecholamines and the related opioid peptide enkephalin within neurons of the rat medulla oblongata. Immunoreactivities for dynorphin and the catecholamine-synthesizing enzyme tyrosine hydroxylase were found to coexist within a limited subpopulation of A2 catecholamine cells, localized to the medial nucleus of the nucleus tractus solitarii. Colocalization of the two opioid peptides was found mainly within perikarya situated in the medial and ventrolateral nuclei of the nucleus tractus solitarii. Triple-labeling studies revealed only rare cases of catecholamine/dynorphin/enkephalin coexistence. These data demonstrate that dynorphin is present within a restricted subpopulation of catecholamine neurons in the dorsal medulla oblongata. In addition, the content of either of the opioids enkephalin or dynorphin appears to distinguish subsets of medullary catecholamine neurons.

Animals↗

Galanin message-associated peptide (GMAP)- and galanin-like immunoreactivities: overlapping and differential distributions in the rat.

Using the indirect immunofluorescence method the distribution of galanin message associated peptide (GMAP)- and galanin-like immunoreactivities (LI) was compared in brain, intestine and some endocrine tissues of rat. In general, neurons in the peripheral and central nervous system contained both immunoreactivities. However, in retina the cones were GMAP-positive but galanin-negative. A strong GMAP-LI was observed in the prolactin cells in the anterior lobe of the pituitary and in the insulin cells in the islets of Langerhans in the pancreas, whereas incubation with galanin antiserum resulted in staining of fewer cells (anterior pituitary) or a very weak fluorescence (pancreas). The results show that most neurons express both GMAP- and galanin-LI, but raise the possibility that in some systems there is a tissue specific, posttranslational differential processing of preprogalanin.

Amino Acid Sequence↗

Evidence for involvement of nitric oxide in the regulation of hypothalamic portal blood flow.

Vasoactive intestinal polypeptide and peptide histidine isoleucine, two peptides with a common precursor and with strong vasodilatory actions, have been suggested to be involved in control of blood flow through the hypothalamic portal blood vessels, in this way regulating the amounts of releasing and inhibitory factors reaching the anterior pituitary. Using the indirect immunofluorescence technique, we now show that this system also contains the enzyme nitric oxide synthase, as well as acetylcholinesterase. It is therefore likely that the control of blood flow through the portal vessels is mediated via relaxation of smooth muscle cells with a high myogenic tone by neuronal release of four vasodilatory compounds, acetylcholine, vasoactive intestinal polypeptide, peptidine histidine isoleucine, and nitric oxide, i.e. a classic neurotransmitter, two neuropeptides and a gas.

Acetylcholine↗

Distribution of thyrotropin-releasing hormone receptor messenger RNA in the rat brain: an in situ hybridization study.

Based on the recent cloning of the mouse thyrotropin-releasing hormone receptor, oligonucleotide probes complementary to the DNA sequence were constructed and used for in situ hybridization studies on the rat brain. Thyrotropin-releasing hormone receptor messenger RNA was found in many areas of the brain, mostly showing high degree of overlap with the distribution thyrotropin-releasing hormone binding sites as previously revealed in autoradiographic studies. Thus, a strong signal was observed in the accessory olfactory bulb, the perirhinal sulcus, the ventral aspects of the hippocampal formation, some amygdaloid nuclei, the diagonal band nucleus, parts of nucleus accumbens, the bed nucleus of the stria terminalis, dorsomedial, lateral and perifornical hypothalamic regions, the septohippocampal nucleus, parts of the vestibular complex, as well as many bulbar motoneurons including the facial, dorsal vagal, ambiguus and hypoglossal nuclei, the superficial layer of the spinal trigeminal nucleus, and motoneurons and dorsal horn neurons in the spinal cord. Cells within one and the same nucleus expressed varying levels of thyrotropin releasing hormone receptor messenger RNA suggesting marked differences in rate of receptor synthesis. Most of these areas receive an input by thyrotropin-releasing hormone-positive nerve endings. Taken together these results suggest that thyrotropin-releasing hormone receptors are mostly localized in the vicinity of the cell bodies which express thyrotropin-releasing hormone receptor messenger RNA and mediate the wide range of actions that have been recorded after administration of exogenous thyrotropin-releasing hormone.

Animals↗