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D Fellmann

Publications and source records attributed to D Fellmann.

At least 19 recordsLinked to original sources

Preprohypocretin (orexin) and prolactin-like immunoreactivity are coexpressed by neurons of the rat lateral hypothalamic area.

Recently, two new neuropeptides named hypocretins I and II (Hcrt) have been described in the rat lateral hypothalamus. The distribution of the neurons expressing these new peptides is similar to that of neurons described in the literature as prolactin (PRL) immunoreactive neurons and specifically labeled by an antiserum (AS) raised against ovine prolactin (oPRL). In this study, we report that every neuron labeled by immunohistochemistry or in situ hybridization for Hcrt is also labeled by the oPRL-AS, and conversely. Furthermore, the labeling of the oPRL-AS is inhibited when this AS is pre-incubated with the 104-109 fragment of the preproHcrt, thus indicating that the oPRL-AS is recognizing an epitope carried by this particular fragment of the preproHcrt. Our previous experimental work on these neurons showed that they may be involved in energy metabolism and water balance, which is in agreement with the current literature about Hcrt functions.

Animals

Prevalence and clinical significance of antiphospholipid antibodies in renal transplant recipients.

BACKGROUND: The prevalence and clinical significance of antiphospholipid antibodies (APAs) have not been extensively studied in non-systemic lupus erythematosus (non-SLE) renal transplant recipients. METHODS: To further define the prevalence and clinical significance of APAs in non-SLE renal transplant recipients and the appearance of dialysis-related APAs after renal transplantation, we conducted a retrospective study on 178 renal transplant recipients. Documentation of anticardiolipin antibodies (ACAs) and lupus anticoagulant in non-SLE renal transplant recipients, retrospective documentation of ACAs on pretransplant frozen plasma and standardized collection of demographic characteristics and posttransplant history of thrombosis were assessed. RESULTS: Fifty of 178 patients (28.1%) had APAs. Transplant duration was shorter and hemodialysis duration was longer in patients with APAs. A posttransplant history of both venous and arterial thrombosis was more frequent in patients with posttransplant APAs (respectively, 18% vs. 6.2% [P<0.001] and 8% vs. 2.3% [P<0.001]). Pretransplant sera were available from 55 patients. Most of patients with posttransplant ACAs had ACAs in the pretransplant period (85%). Pretransplant ACAs were associated with a posttransplant history of venous thrombosis (P<0.001). CONCLUSIONS: Our study demonstrates a high prevalence of APAs in non-SLE renal transplant recipients. Most of them have been acquired in the pretransplant period. Both pretransplant ACAs and posttransplant APAs are associated with posttransplant episodes of thrombosis. Further studies are required to determine the interest of prophylactic measures.

Adult

Polyethylene glycol-induced hypovolemia affects the expression of MCH mRNA, but not dynorphin or secretogranin II mRNAs, in the rat lateral hypothalamus.

Two prominent neuron populations of the rat lateral hypothalamus express genes encoding respectively the prepromelanin-concentrating hormone (MCH) or dynorphin (DYN) and secretogranin II (SGII). Their roles remain hypothetical in mammals. In the present study, we examined the changes in MCH, DYN and SGII gene expression in dehydrated rats compared to controls. Dehydration was obtained by subcutaneous injection of polyethylene glycol (PEG) resulting in a large reduction of the extracellular fluid volume. Using competitive semi-quantitative RT-PCR and in situ hybridization methods, PEG-injected animals showed a significant increase of MCH mRNA level but no variation of DYN and SGII mRNA levels. These results confirm previous observations suggesting that intra- and extracellular dehydration challenges affect different regulation circuits; they indicate that both neuron populations could be involved in the maintenance of body fluid homeostasis, directly, or indirectly, as integrators of various information leading to goal-oriented behaviour.

Animals

Beta-adrenergic and muscarinic receptor mRNA accumulation in the sinoatrial node area of adult and senescent rat hearts.

The sinoatrial (SA) node is the cardiac pacemaker and changes in its adrenergic-muscarinic phenotype have been postulated as a determinant of age-associated modifications in heart rate variability. To address this question, right atria were microdissected, the SA node area was identified by acetylcholinesterase staining, and, using a RT-PCR method, the accumulation of mRNA molecules encoding beta1- and beta2-adrenergic (beta1- and beta2-AR) and muscarinic (M2-R) receptor was quantified to define the proportion between beta-AR and M2-R mRNAs within the sinoatrial area of adult (3 months) and senescent (24 months) individual rat hearts. In adult hearts, the highest M2-R/beta-AR mRNA ratio was observed within the sinoatrial area compared with adjacent atrial myocardium, while in the senescent hearts, no difference was observed between sinoatrial and adjacent areas. This change was specific of the sinoatrial area since adult and senescent whole atrial or ventricular myocardium did not differ in their M2-R/beta-AR mRNA ratio, and was associated with a fragmentation of acetylcholinesterase staining of the senescent SA node. Quantitative changes in the expression of genes encoding proteins involved in heart rate regulation specifically affect the sinoatrial area of the senescent heart.

Aging

Localized deposition of M-cadherin in the glomeruli of the granular layer during the postnatal development of mouse cerebellum.

M-cadherin is a Ca2+-dependent cell adhesion molecule of the cadherin family, initially localized at the areas of contact between myotubes during myogenesis, but also detected in the peripheral nerve and at the adult neuromuscular junction. In this study, searching for the expression of M-cadherin in the adult mouse brain, we observed a restricted expression of M-cadherin in one of the three layers of the cerebellar cortex: the granular layer. M-cadherin was accumulated in structures rich in synapses and other intercellular junctions where mossy fibers connect granule cell dendrites, the glomeruli. This molecule was not expressed in the cerebellum during the first steps of postnatal cerebellar neurogenesis: granule cell proliferation and migration and Purkinje cell alignment. M-cadherin expression was first detected at postnatal day (P) 11, after the establishment of the synaptic connections between mossy fibers and granule cell dendrites. It then accumulated in glomeruli during their phase of maturation which is characterized by the formation of puncta adherentia between granule cell dendrites. M-cadherin was undetectable in the cerebella of the weaver and staggerer mutants, lacking granule cells, and therefore mature glomeruli and puncta adherentia. Furthermore, other components classically associated with intercellular junctions, i.e., alpha-caterin, beta-catenin and actin filaments, closely paralleled M-cadherin appearance and colocalized with M-cadherin in the mature glomeruli. M-cadherin, which appears as a molecular marker of glomerulus maturation, might be implicated in the formation, and be the ligand, of adherens junctions encountered in this structure.

Actins

Changes in rat melanin-concentrating hormone and dynorphin messenger ribonucleic acids induced by food deprivation.

Melanin-concentrating hormone (MCH) and dynorphin genes are expressed in two discrete neuron populations of the rat lateral hypothalamus. Their roles remain hypothetical in mammals. In order to analyse changes in MCH and dynorphin gene expression, a multiplex competitive semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was developed to assay their mRNAs. This technique was used to examine MCH and dynorphin mRNA content in 24-h and 48-h food-deprived rats compared to controls. A two-fold induction of dynorphin mRNA by 24 h, followed by a sharp decrease at 48 h were observed. A moderate increase in MCH mRNA content was noticed by 24 h; 48 h of fasting restored the control levels.

Animals

Exploring the expression of the melanin-concentrating hormone messenger RNA in the rat lateral hypothalamus after goldthioglucose injection.

Melanin-concentrating hormone (MCH) is expressed in a large neuronal population of the rat lateral hypothalamus. This area is known to be implicated in the regulation of thirst and hunger and to contain glucose-sensitive cells. In the present study, we investigated the effects of goldthioglucose (GTG), a toxic form of glucose, on the expression of the MCH gene in the rat lateral hypothalamus by immunocytochemistry, in situ hybridization and competitive RT-PCR. We observed that the MCH immunoreactivity and the level of MCH mRNA were not changed after intraperitoneal GTG injection (0.35 mg/g body weight). These results together with previous data suggest that the glucose-sensitive cells of the lateral hypothalamus are different from the MCH neurons and remain to be identified.

Animals

Immunocytochemical detection of the neurokinin B receptor (NK3) on melanin-concentrating hormone (MCH) neurons in rat brain.

The presence of the neurokinin B receptor (NK3 receptor) in the rat lateral hypothalamus and the zona incerta was previously reported. The aim of the present study was to define its cellular localization in these areas. Investigations, coupling immunocytochemical and in situ hybridization techniques, focussed on two neuron populations: the melanin-concentrating hormone (MCH) neurons and a population of neurons recognized by an ovine prolactin antiserum (PRL-ir neurons). While PRL-ir neurons did not exhibit NK3 immunoreactivity, 57% +/- 6% of MCH neurons were strongly stained by the NK3 antiserum. These results suggest that neurokinin B is involved in the regulation of MCH neuron activity via the NK3 receptor; they provide new bases for further investigations on MCH role in the control of food and water intake.

Animals

[Study of gene expression of melanin-concentrating hormone and of dynorphin in the rat hypothalamus by multiplex competitive RT-PCR].

Melanin-concentrating hormone (MCH) and dynorphin genes are expressed in two discrete neuron populations of the rat lateral hypothalamus. Their roles remain hypothetical in mammals. In order to analyze changes in MCH and dynorphin gene expression, we developed a multiplex competitive semi-quantitative RT-PCR allowing to study simultaneously the variations of their mRNAs. This technique was used to examine MCH and dynorphin mRNAs contents in rats food-deprived for 24 or 48 hours as compared to controls. A 2-fold induction of dynorphin mRNA by 24 hours, followed by a sharp decrease at 48 hours and return to control level were observed. A moderate increase in MCH mRNA content was noticed by 24 hours. A 48 hours fasting restored the control levels.

Animals

Alteration of dynorphin and secretogranin II in the prolactin immunoreactive neurons of the rat lateral hypothalamus upon osmotic stimulation.

The prolactin immunoreactive neurons of the rat lateral hypothalamus were previously reported to express the dynorphin and secretogranin II genes. In the present study, the response of these neurons to osmotic challenge was immunocytologically investigated by using prolactin, dynorphin, secretogranin II and c-Fos antisera. In addition, the mRNA levels for secretogranin II and dynorphin were compared by in situ hybridization in controls and salt-loaded rats. For this model of chronic hyperosmolality, the prolactin and c-Fos immunoreactivities were not stimulated by salt drinking, but dynorphin and secretogranin II immunoreactivities as well as mRNA levels for dynorphin and secretogranin II significantly increased in the lateral hypothalamus. We suggest that the prolactin-immunoreactive neurons may be involved in the regulation of water homeostasis.

Animals

Distribution of melanin-concentrating hormone (MCH)-like immunoreactivity in neurons of the diencephalon of sheep.

An immunohistochemical study with an antiserum raised against salmon melanin concentrating-hormone has demonstrated the presence of numerous melanin concentrating-hormone-immunoreactive neurons in the lateral hypothalamic areas of the sheep. The pattern of distribution of these perikarya is similar to that of rodents and primates. In sheep, however, melanin concentrating-hormone-immunoreactive neurons appeared to form two gatherings: the first is situated ventromedially to the internal capsule and the second in the dorsolateral hypothalamus. In these areas, numerous immunostained perikarya are observed. Compared to the rats, labelled neurons extended more caudally in the ventral tegmental area and more rostrally above the optic chiasma. Compared to primates, these neurons are less numerous in the periventricular area. In our study, dense networks of melanin concentrating-hormone-immunoreactive varicose fibers were observed in the supramamillary nucleus, the lateral hypothalamus, the nucleus medialis thalami and nucleus reuniens and in the bed nucleus of the stria terminalis.

Animals

The synthesis of melanin-concentrating hormone is stimulated by ventromedial hypothalamic lesions in the rat lateral hypothalamus: a time-course study.

The activity of melanin-concentrating (MCH) neurons, was investigated by immunocytochemical and hybridocytochemical techniques in male rats bearing limited lesions of the ventromedial hypothalamic nuclei (VMN). 2 days after operation, the abundance of immunoreactive cell bodies and fibres and the intensity of labelling seemed slightly decreased in lesioned rats as compared to controls while no significant difference could be detected in MCH gene expression. After 8 days, synthesis, storage and transport of MCH appeared strongly stimulated and this stimulation lasted until the end of the experiment (day 35), suggesting that VMN plays a physiological role in controlling MCH neuron activity.

Animals

Occurrence of secretogranin II in the prolactin-immunoreactive neurons of the rat lateral hypothalamus: an in situ hybridization and immunocytochemical study.

The occurrence of secretogranin II in a neuron population of the rat lateral hypothalamus specifically detected by an anti-serum to ovine prolactin was examined. As this population was previously reported to synthesize dynorphin, the distribution of neurons recognized by ovine prolactin-, dynorphin B- and secretogranin II anti-sera was investigated on adjacent sections of hypothalami. The prolactin immunoreactive neurons were the only cells in the lateral hypothalamus to be stained by secretogranin II anti-serum. Moreover, coupling immunocytochemical detection and in situ hybridization with an oligonucleotide probe complementary to secretogranin II mRNA showed that these neurons expressed the secretogranin II gene. These new findings should help to study the physiological role of the prolactin immunoreactive neurons of the lateral hypothalamus.

Animals

Early detection of secretogranin-II (SgII) in the human fetal pituitary: immunocytochemical study using an antiserum raised against a human recombinant SgII.

Secretogranin-II (SgII) is a protein contained within secretory granules of mainly gonadotrophs. The purpose of this study was to determine whether SgII immunoreactivity (SgII-IR) in the human fetal pituitary was temporally related to gonadotropin immunoreactivity. A specific antihuman SgII antiserum was thus required. A complementary DNA clone with an open reading frame for human (h) SgII was synthesized by reverse transcription-polymerase chain reaction from pituitary total RNA. This clone was used to obtain the SgII polypeptide (-9 to 152) as a fusion protein, in a heterologous expression prokaryotic system. Antisera against the fusion protein were raised in rabbits and checked for specificity and sensitivity through Western blotting. Human fetal pituitaries from week 6 of gestation onward were used for immunocytochemical studies. Consecutive semithin sections were treated with the specific antisera against hSgII, beta-endorphin, and hPRL and with monoclonal antibodies to hCG alpha, hLH, and hFSH. SgII immunoreactivity appeared at week 8 and was restricted to pituitary cells expressing beta-endorphin (100% colocalization). At week 9, FSH-positive cells did not contain SgII. From week 10, gonadotrophs progressively exhibited SgII-IR, up to 50% of that in FSH-containing cells at week 26. The granin was never found in PRL cells whatever the stage of development. The present data demonstrate that SgII-IR is detected very early in fetal life; however, the positive cells are not gonadotrophs, but corticotrophs. Within gonadotrophs, SgII appears subsequent to hormones. At birth, more than 90% of SgII-IR cells are represented by corticotrophs and gonadotrophs.

Adrenal Glands

Induction of Fos-like immunoreactivity in rat oxytocin neurons following insulin injections.

Double immunostaining for oxytocin (OT) and Fos was used to study the oxytocinergic system of the rat hypothalamic paraventricular nucleus (PVN) following intraperitoneal insulin injections. The expression of c-fos in the PVN appeared about 3 h after insulin treatment and was very high after 5 h while no labelling was observed in isotonic saline-injected animals. Twelve to 18% of OT neurons expressed Fos-like immunoreactivity and these activated neurons were found in both the magno- and the parvocellular compartments of the PVN suggesting that the OT neuron responses to insulin induced disturbances are complex and involve hormonal as well as autonomic pathways.

Animals