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F Nürnberger

Publications and source records attributed to F Nürnberger.

At least 19 recordsLinked to original sources
PubMed2002

Transcription factor CREB and its stimulus-dependent phosphorylation in cell and explant cultures of the bovine subcommissural organ.

The subcommissural organ (SCO) is an ependymal brain gland that synthesizes and secretes glycoproteins. Very little is known about the signal transduction cascades operating in this organ and their impact on gene expression. An important transcription factor that regulates gene expression in glial cells and neurons is the cyclic-AMP-responsive element binding protein (CREB), which is activated by phosphorylation of the serine residue 133. Here, we analyzed the presence of CREB in bovine SCO cells and its phosphorylation by drugs that activate cyclic-AMP-dependent or calcium-dependent signal transduction pathways. We also investigated the effects of three natural signaling molecules, serotonin (5HT), substance P (SP) and ATP, on CREB phosphorylation and on the second messengers cyclic AMP and calcium. Investigations were performed with cell and explant cultures by using immunocytochemistry, immunoblot, enzyme-linked immunosorbent assay, and the Fura-2 technique. A strong immunosignal for total (phosphorylated and unphosphorylated) CREB was found in virtually all SCO cells. Total CREB levels did not change upon stimulation. Phosphorylated (p)CREB levels were low in unstimulated cells and significantly elevated by drugs that increase the levels of cyclic AMP or free calcium ions. pCREB was also induced by SP and ATP; both substances increased the intracellular calcium concentration but did not affect the formation of intracellular cyclic AMP. 5HT did not influence the phosphorylation of CREB, the intracellular calcium concentration, or the formation of cyclic AMP. Our data identify CREB as an SCO transcription factor that can be activated by the second messengers cAMP and calcium. SP and ATP stimulate the phosphorylation of CREB apparently via a calcium-dependent mechanism and are thus involved in the control of gene expression in the bovine SCO.

Animals↗
PubMed2001

Effects of neuroactive substances on the activity of subcommissural organ cells in dispersed cell and explant cultures.

The subcommissural organ (SCO), an ependymal (glial) circumventricular organ, releases glycoproteins into the cerebrospinal fluid; however, the regulation of its secretory activity is largely unknown. To identify neuroactive substances that may regulate SCO activity, we investigated immunocytochemically identified bovine SCO cells by means of calcium imaging. This analysis was focused on: (1) serotonin (5HT) and substance P (SP), immunocytochemically shown to be present in axons innervating the bovine SCO; and (2) ATP, known to activate glial cells. 5HT had no effect on the intracellular calcium concentration ([Ca(2+)](i)), and its precise role remains to be clarified. SP elicited rises in [Ca(2+)](i) in approx. 30% and ATP in even 85% of the analyzed SCO cells. These effects were dose-dependent, involved NK(3) and P2Y(2) receptors linked to G protein and phospholipase C (PLC) activation, and could not be mimicked by forskolin or 8-bromo-cAMP. In 50% of the SP-sensitive cells, the increases in [Ca(2+)](i) comprised calcium release from thapsigargin-sensitive intracellular stores and an influx of extracellular calcium via protein kinase C (PKC)-induced opening of L-type voltage-gated calcium channels (VGCCs). In the remaining SP-sensitive cells, the increase in [Ca(2+)](i) was caused exclusively by influx of extracellular calcium via VGCCs of the L-type. In all ATP-sensitive cells the increase in [Ca(2+)](i) involved calcium release from thapsigargin-sensitive intracellular stores and a PKC-mediated influx of extracellular calcium via L-type VGCCs. Our data suggest that SP and ATP are involved in regulation of the activity of SCO cells.

Adenosine Triphosphate↗
PubMed2001

The dispersed cell culture as model for functional studies of the subcommissural organ: preparation and characterization of the culture system.

The subcommissural organ (SCO) is an enigmatic secretory gland of the brain, which is believed to be derived from ependymal (glial) precursor cells. We here developed a dispersed cell culture system of the bovine SCO as an approach to functional analyses of this brain gland. Tissue of the bovine SCO obtained from the slaughterhouse was papain dissociated either directly after dissection or after preparation of SCO explants. The latter had been maintained for 4-6 weeks in organ culture. The dispersed cells were cultured for up to 14 days and continuously tested for their secretory state by immunostaining of their secretory product. With respect to the morphology of the SCO cells (shape, processes, nucleus), no difference was found between the culture of freshly dissociated SCOs and that of dissociated SCO explants. In all cases, the dissociation caused a dedifferentiation; typical elongated cells were formed increasingly after 1 day of culture. Thereafter, only the cellular size increased, whereas the shape and the viability of the cells remained unchanged. Proliferating SCO cells were never observed. The culture obtained from fresh SCO tissue contained more glia cells and fibrocytes than the culture prepared from SCO explants. The proliferation of glia cells and fibrocytes was suppressed by blocking the mitotic activity with cytosine-beta-D-arabino furanoside (CAF). The cytophysiological features of the cultured dispersed cells of both origins did not differ as demonstrated by classical histology, by immunocytochemistry for the secretory products of the SCO, by the characteristics of calcium influx into the cytoplasm ([Ca2+]i) and cyclic adenosine monophosphate (cAMP) after stimulation with adenosine-5-triphosphate, substance P or serotonin, and by the activation of the transcription factor cAMP-responsive element-binding protein. Because of the maintenance of their viability, their capacity to release the secretory product into the culture medium, their receptive capacity, and their signal transduction pathways, we conclude that the dispersed cell culture system, especially that obtained from SCO explants, represents an appropriate and useful model for functional studies of the mammalian SCO.

Adenosine Triphosphate↗
PubMed2001

Presence and functional significance of neuropeptide and neurotransmitter receptors in subcommissural organ cells.

The subcommissural organ (SCO) of mammals is innervated by several neuropeptide and neurotransmitter systems. So far, substance P (SP), oxytocin (OXT), vasopressin (VP), somatostatin (SOM), thyrotropin-releasing factor (TRF), and angiotensin II (ANGII) were identified in neuropeptidergic input systems, and serotonin (5HT), gamma-amino butyric acid (GABA), noradrenaline (NA), dopamine (DA), and acetylcholine (Ach) were neurotransmitters observed in systems afferent to the SCO. In the present report, based on literature data and our own investigations, we describe the occurrence of peptide and transmitter receptors in the SCO by means of autoradiographic and biochemical studies. Further, we summarize aspects of the signal transduction cascades possibly linked to different receptor types of the SCO; these studies included the use of calcium imaging (FURA-2 technique), ELISA technique, and immunocytochemistry. Receptors were identified for adenosine, angiotensin II, imidazoline, glucocorticoids, mineralocorticoids, NA, and embryonic brain kinase. The studies on intracellular signal-transduction indicated receptors for tachykinins and for ATP. In SCO cells, Ca(++) and c-AMP were identified to act as second messengers. As important transcription factor, cAMP-/Ca(++)-response element binding protein (CREB) was observed. Ach and NA did not show a significant effect on the subcommissural signal transduction.

Adenosine↗
PubMed1998

Dynamic changes in the immunoreactivity of neuropeptide systems of the suprachiasmatic nuclei in golden hamsters during the sleep-wake cycle.

The sleep-wake cycle is virtually the most prominent circadian rhythm in mammals. In the timing system of sleep and wakefulness, the intrinsic neuropeptide systems of the suprachiasmatic nuclei (SCN) may play an important role. To elucidate this possible influence in the golden hamster, the immunoreactivity patterns of the suprachiasmatic gastrin-releasing peptide (GRP), vasoactive intestinal polypeptide (VIP), and arginine-vasopressin (AVP) systems were investigated in relation to the day-night and sleep-wake cycle by use of immunocytochemistry combined with semiquantitative planimetric analysis. For the GRP system, the highest level of immunoreactivity (expressed as area density) was observed in sleeping hamsters. Intermediate levels were found in awake, motorically active evening animals, whereas the lowest levels of immunoreactivity were detected in awake, motorically inactive hamsters studied in the morning. The immunoreactivity of the VIP system showed a completely opposite pattern, indicating highest area density in awake morning, intermediate area density in awake evening and lowest area density in sleeping golden hamsters. The immunoreactivity pattern of the AVP system, displaying highest levels in sleeping individuals, was virtually identical to that of the GRP system. Together with the related signs of neuronal activity, the present results favor an important role of these neuropeptide systems for the integration of central nervous information related to the sleep-wake cycle with photic information of the retinal input.

Animals↗
PubMed1997

The somatostatin system of the brain and hibernation in the European hamster (Cricetus cricetus).

The depression of physiological processes characteristic of mammalian hibernation is precisely regulated by the central nervous system, especially by the neuropeptidergic apparatus of the hypothalamus. Because of inhibitory influences on neuronal circuits within the brain and suppressive effects on the metabolism via the endocrine axis, somatostatin has been implicated in the regulation of hibernation. The somatostatin system of the brain was investigated with immunocytochemistry, in situ hybridization, and radioimmunoassays in euthermic summer, euthermic winter, and hibernating European hamsters (Cricetus cricetus). Numerous somatostatin-immunoreactive perikarya were observed in the periventricular hypothalamic nucleus. The striatum, amygdala, and cortex contained only scattered immunoreactive perikarya. These entities also contained immunoreactive fiber profiles, although the highest density of immunoreactive fibers was found in the median eminence. Immunocytochemistry and radioimmunoassays showed that the number of somatostatin-immunoreactive perikarya and fibers and the content of somatostatin in the hypothalamus and the median eminence was conspicuously lower in euthermic winter animals than in euthermic summer animals. This decrease was more pronounced in hibernating specimens. In situ hybridization also demonstrated a decrease in the expression and synthesis rate of somatostatin in euthermic winter animals; again, this was even more dramatic in hibernating hamsters. These changes were less pronounced or non-significant in the extrahypothalamic somatostatin-immunoreactive perikarya and fiber systems, as shown by immunocytochemistry and radioimmunoassay, respectively.

Animals↗
PubMed1997

Beta-adrenergic signal transduction in the hypothalamus of the European hamster: relation with the seasonal hibernation cycle and the diurnal activity cycle.

Mammalian hibernation, an adaptation to survive harsh winter conditions, is one of the most prominent seasonal rhythmic processes exactly regulated on a low metabolic level. Diurnal variations in vegetative physiology are missing during hibernation; however, a precisely working diurnal system is mandatory for both the proper initiation and termination of the annual hibernation phase and the periodical arousal reactions. Biorhythms and the vegetative physiological processes connected with hibernation are, among others, controlled by hypothalamic noradrenaline systems. In this study, the density, binding capacity, and relative proportions of beta 1- and beta 2-adrenergic receptors (AR) within the hypothalamus of: 1) motorically inactive summer; 2) motorically active summer; 3) aroused, motorically active winter; and 4) deeply hibernating winter European hamsters (Cricetus cricetus) were studied. For further analysis of the beta-adrenergic signal transduction cascade, the activity of adenylyl cyclase (AC) was measured by formation of cAMP in controls, after stimulation of G proteins, or after forskolin stimulation without or in presence of manganese ions. While beta 1- and beta 2-AR subtypes were nearly equally abundant (50% beta 1:50% beta 2) in active summer, inactive summer, and hibernating hamsters, a significant redistribution in favor of beta 2-AR occurred after arousal (40% beta 1:60% beta 2). The activity of AC was much higher in active summer hamsters than in inactive summer, aroused winter, and hibernating winter hamsters. When AC was stimulated by guanylylimidophosphate [Gpp(NH)p], MnCl2, forskolin, or by forskolin in presence of MnCl2 instead of MgCl2, the potency to stimulate AC was found to show the following rank order: basal < Gpp(NH)p < MnCl2 < or = forskolin + MnCl2 < forskolin.

1-Methyl-3-isobutylxanthine↗
PubMed1995

Somatostatin-immunoreactive cell in the gastrointestinal tract of the frog Rana esculenta.

The morphology and topographic distribution of somatostatin-immunoreactive cells in the stomach and small intestine of the frog Rana esculenta were studied at the light-microscopic level by the use of the peroxidase-antiperoxidase method. Scattered immunostained cells occurred in all regions of the gastrointestinal tract investigated. In the small intestine, the number of these cells decreased gradually in the oral to anal direction, i.e. from the pyloric (antral) stomach to the entrance into the colon. Most of the immunostained cells possessed thick, short cytoplasmic processes, which did not display a preferential spatial orientation. Other somatostatin-immunoreactive cells, which were exclusively located in the small intestine, gave rise to a single long extension oriented toward the lumen. In both stomach and small intestine, a complete penetration of the epithelial surface by these processes of somatostatin-immunoreactive cells was observed only occasionally. The morphological features of the somatostatin-immunostained cells speak in favor of endocrine, paracrine, and possibly also intraluminal secretory functions of the enteroendocrine somatostatin system in frogs.

Animals↗
PubMed1995

The neuroendocrine system in hibernating mammals: present knowledge and open questions.

The present review describes the distribution and the function-dependent reactivity pattern of those peptidergic and aminergic components of the neuroendocrine system of hibernating mammals that have been studied by histological, pharmacological and physiological techniques. Particular attention has been paid to the intrinsic connectivity of the peptidergic apparatus and its input systems. Since the reactivity patterns of the neuroendocrine system show remarkable fluctuations in relation to the various stages of hibernation and euthermia, these fluctuations have been analyzed with respect to (1) their causative role in the regulation of hibernation and (2) their secondary response to physiological changes during hibernation. The author's investigations described in this review have mainly been performed in European hedgehogs (Erinaceus europaeus), European and golden hamsters (Cricetus cricetus, Mesocricetus auratus), dormice (Glis glis), and in Richardson's and Columbian ground squirrels (Spermophilus richardsonii, Spermophilus columbianus), by the use of light- and electron-microscopic immunocytochemistry and histochemistry, in situ hybridization, radioimmunoassays and stereotaxically guided application techniques. These experiments were also performed in hypothermic animals. The (partially published) results obtained by the author and his associates are reviewed with reference to the body of evidence found in the recent literature. With respect to their reactivity patterns, several neuropeptide and transmitter systems can be regarded as candidates for control systems of hibernation. Neuronal complexes immunoreactive for endogenous opiates, in particular enkephalin, and also for vasopressin, somatostatin, substance P, corticotropin-releasing factor and serotonin are probably involved in the neuroendocrine control of hibernation.

Animals↗
PubMed1995

Distribution and ultrastructure of somatostatin-immunoreactive cells in the pancreas of Rana esculenta.

Apart from a description of the general organization of the endocrine pancreas, the present study is focussed on the distribution and ultrastructural morphology of somatostatin-immunoreactive cells in the pancreas of the frog Rana esculenta. For light-microscopic histochemistry, the peroxidase-antiperoxidase technique was used. For the ultrastructural investigation, we employed the immunogold method. The endocrine pancreas of R. esculenta is composed of numerous islet-like structures, which contain several small somatostatin-immunoreactive cells arranged in the form of clusters. Often, however, single somatostatin cells are randomly distributed among the acinar tissue of the pancreas. These individually arranged elements possess long processes which terminate on exocrine pancreatic cells. The ultrastructural features of somatostatin-immunoreactive cells speak in favor of their endocrine and paracrine functions.

Animals↗
PubMed1994

Effects of illumination and enucleation on substance-P-immunoreactive structures in subcortical visual centers of golden hamster and Wistar rat.

The undecapeptide substance P is found in different entities of the visual system that control eye movement and synchronize endogenous rhythms with the light cycle (i.e., superior colliculus, suprachiasmatic nucleus, intergeniculate leaflet). Immunocytochemical methods were used to compare the reactivity to substance P in the brain of five groups of golden hamsters and two groups of Wistar rats: (1) untreated hamsters kept under 14L:10D and sacrificed at noon; (2) identically maintained animals sacrificed at midnight; (3) enucleated animals kept under control conditions; (4) hamsters kept under constant darkness; (5) hamsters kept under the same conditions as the controls, but intraventricularly injected with colchicine. The results obtained in golden hamsters of groups (1) and (3) were compared with findings in Wistar rats treated accordingly [groups (6) and (7)]. Substance P-immunoreactive perikarya were found in the suprachiasmatic nucleus and superior colliculus of hamsters and Wistar rats. Substance P-immunoreactive nerve fibers were abundant in the hypothalamic area ventral to the paraventricular nucleus, in the intergeniculate leaflet, in some thalamic nuclei, and in the superior colliculus. Immunoreactivity to substance P in the suprachiasmatic nucleus and intergeniculate leaflet did not vary among the experimental groups. However, a conspicuous decrease in reactivity to substance P was observed in the superficial layers of the superior colliculus of enucleated hamsters and rats, compared with all other groups. These results indicate that substance P immunoreactivity in the superior colliculus, but not that in the suprachiasmatic nucleus or intergeniculate leaflet, depends on the integrity of the retinal projection.

Animals↗
PubMed1993

Acute monoblastic leukemia with skin nodules in an adult.

An 18-year-old man had a 3-week history of malaise, a 1-week history of a papular eruption, and increasingly severe multisystem neurologic symptoms. A diagnosis of acute monoblastic leukemia was made. Immunophenotypic characterization of peripheral blood, bone marrow, and cutaneous infiltrates revealed a predominant myelomonocytic phenotype with the coexpression of intercellular adhesion molecule type 1, IgE receptor, terminal deoxynucleotidyl transferase, and some T-cell markers. These findings may have important clinical and pathogenetic implications regarding the biologic and pathologic behavior of the myelomonocytic leukemic cells. They also explain in part the impressive presentation of the disease in the skin.

Adolescent↗
PubMed1993

[Treatment of vitiligo with phenylalanine and UV-A].

Seven patients with generalized vitiligo were treated with 100 mg/kg body weight phenylalanine in combination with UV-A. A 20% repigmentation of the vitiligenous area was achieved in only one patient. Three patients each showed either a sparse follicular repigmentation or no effect at all. Because of the disappointing therapeutic results and possible side effects, we cannot recommend a therapy with phenylalanine and UV-A for vitiligo.

Administration, Oral↗
PubMed1991

The efferent connections of the lateral septal nucleus in the guinea pig: projections to the diencephalon and brainstem.

The anterograde Phaseolus vulgaris-leucoagglutinin (PHA-L) tracing technique was used to determine the distribution of efferent fibers originating in the lateral septal nucleus of the guinea pig. For complementary detection of the chemical identity of the target neurons, double-labeling immunocytochemistry was performed with antibodies to PHA-L and to vasopressin, oxytocin, vasoactive intestinal polypeptide, serotonin or dopamine beta-hydroxylase, respectively. The hypothalamus received the majority of the PHA-L-stained septofugal fibers. Here, a specific topography was observed. (1) The medial and lateral preoptic area, (2) the anterior, lateral, dorsal, posterior hypothalamic and retrochiasmatic area, (3) the supraoptic, paraventricular, suprachiasmatic, dorsomedial, caudal ventromedial and arcuate nuclei, and (4) the tuberomammillary, medial and lateral supramammillary, dorsal and ventral premammillary nuclei always contained PHA-L-labeled fibers. The rostral portion of the ventromedial nucleus and the medial and lateral mammillary nucleus only occasionally showed weak terminal labeling. In other diencephalic areas, termination of PHA-L-labeled fibers was observed in the epithalamus and the nuclei of the midline region of the thalamus. In the mesencephalon, terminal varicosities occurred in the ventral tegmental area, interfascicular and interpeduncular nucleus, and periaqueductal gray. In addition, the dorsal and medial raphe nuclei of the metencephalon, together with the locus coeruleus and the dorsal tegmental nucleus, received lateral septal efferents.

Animals↗
PubMed1991

The efferent connections of the lateral septal nucleus in the guinea pig: intrinsic connectivity of the septum and projections to other telencephalic areas.

The distribution of efferent fibers originating in the lateral septal nucleus was investigated in guinea pigs by means of anterograde tracing with Phaseolus vulgaris-leucoagglutinin (PHA-L). Special emphasis was placed on the intraseptal fiber systems. The fibers originating from the different subnuclei of the lateral septal nucleus formed massive horizontal connections in the rostrocaudal axis. Projections to the contralateral, congruent subnuclei were also detected. In the medial septum/diagonal band of Broca complex the largest number of PHA-L-stained fibers was found after application of the tracer into the dorsal subnucleus of the lateral septal nucleus; the density of the efferent fibers decreased progressively after injection into the intermediate or ventral subnuclei. In all cases the diagonal band contained a much higher number of efferent fibers from the lateral septal nucleus than from the medial septal nucleus. In the medial septal nucleus, terminal labeling was generally sparse. Other telencephalic areas (organum vasculosum of the lamina terminalis, nucleus accumbens, bed nucleus of the stria terminalis, amygdala, hippocampal complex, and other cortical areas) contained varying numbers of labeled projections. In double-labeling experiments, a close spatial relationship between PHA-L-stained fibers and vasoactive intestinal polypeptide-immunoreactive perikarya was observed in several of these target areas.

Animals↗