Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 175
Filter
1.
J Neuroendocrinol ; 29(12)2017 12.
Article in English | MEDLINE | ID: mdl-29024103

ABSTRACT

Interleukin (IL)-6- /- mice develop mature onset obesity, whereas i.c.v. injection of IL-6 decreases obesity in rodents. Moreover, levels of IL-6 in cerebrospinal fluid (CSF) were reported to be inversely correlated with obesity in humans. Tanycytes lining the base of the third ventricle (3V) in the hypothalamus have recently been reported to be of importance for metabolism. In the present study, we investigated whether tanycytes could respond to IL-6 in the CSF. With immunohistochemistry using a well characterised antibody directed against the ligand binding receptor for IL-6, IL-6 receptor α (IL-6Rα), it was found that tanycytes, identified by the two markers, vimentin and dopamine- and cAMP-regulated phosphoprotein of 32 kDa, contained IL-6Rα. There were fewer IL-6Rα on another type of ventricle-lining cells, ependymal cells, as identified by the marker glucose transporter-1. To demonstrate that the immunoreactive IL-6Rα were responsive to IL-6, we injected IL-6 i.c.v. This treatment increased immunoreactive phosphorylated signal transducer and activator of transcription-3 (pSTAT3) in tanycytes after 5 minutes and in cells in the medial part of the arcuate nucleus after 5 and 15 minutes. Intracerebroventricular injection of leptin exerted similar effects. As expected, i.p. injection of leptin also induced pSTAT3 staining in the hypothalamus, whereas i.p. IL-6 injection had little effect on this parameter. Intracerebroventricular or i.p. injection of vehicle only had no effect on pSTAT3-immunoreactivity. In summary, there are functional IL-6Rα on tanycytes at the bottom of the 3V, in agreement with the possibility that ventricular administration of IL-6 decreases obesity in mice via an effect on this cell type.


Subject(s)
Ependymoglial Cells/metabolism , Interleukin-6 Receptor alpha Subunit/metabolism , Third Ventricle/cytology , Animals , Arcuate Nucleus of Hypothalamus/metabolism , Ependymoglial Cells/cytology , Female , Leptin/administration & dosage , Leptin/metabolism , Male , Mice, Inbred C57BL , Mice, Knockout , Phosphorylation , STAT3 Transcription Factor/metabolism , Signal Transduction
2.
World J Urol ; 33(7): 1039-43, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25164043

ABSTRACT

BACKGROUND AND PURPOSE: Residual calculi after stone therapy need to be treated if they are clinically significant, mainly depending on the size of the calculi. There are different ways to detect and measure the size of residual calculi as for example KUB or computed tomography. The Uro Dyna-CT (Siemens Healthcare solutions, Erlangen, Germany) allows cross-sectional imaging and 3D reconstructions during endourological interventions. In this ex vivo study, we investigate the accuracy of imaging residual calculi with the Uro Dyna-CT. MATERIALS AND METHODS: Twenty-seven artificial stones (plaster of Paris) were scanned with the Uro Dyna-CT by a special urolithiasis protocol and post-image processing with 3D-reconstruction and cross-sectional imaging was performed. The major diameter of each stone was measured at the dedicated workstation by one investigator. The same stones were measured randomized with a digital caliper (real size). Finally, the two measurements were compared. The paired t test, Pearson's correlation coefficient, the F Test, a reference area for differences, the intraclass correlation coefficient, the Maloney-Rastogi test and the Bland-Altman analysis were used for statistical analyses. RESULTS: The range of stone sizes was 3-5 mm. We did not find significant differences in the size of the stones measured with the Uro Dyna-CT and the digital caliper (paired t test p = 0.3597) and we showed a good correlation between the two measuring methods (intraclass correlation coefficient 0.4465; p = 0.0088). CONCLUSION: Renal calculi can be measured highly accurately with the Uro Dyna-CT. Whether this technique will lead to less residual fragments after stone treatment needs to be shown in further studies.


Subject(s)
Imaging, Three-Dimensional , Tomography, X-Ray Computed , Urolithiasis/diagnosis , Humans , Models, Biological , Reproducibility of Results
3.
Klin Padiatr ; 225(3): 159-63, 2013 May.
Article in English | MEDLINE | ID: mdl-23625682

ABSTRACT

BACKGROUND: Anemia in toddlers may result from many disorders including excessive feeding with cow's milk. Another sequel of age-inadequate cow's milk nutrition may be gastric lactobezoar (GLB), a dense lump of coagulated milk and mucus in the stomach. PATIENTS: 3 toddlers presented with a history of excessive intake of full cream cow's milk, abdominal distension, vomiting, dehydration, fatigue, marked pallor and tachycardia. DIAGNOSTIC WORKUP: Diagnostic imaging revea-led large GLBs as the likely origin of the abdominal symptoms. Laboratory evaluation showed severe anemia with depleted iron stores and signs of protein catabolism. Non-cow's milk-induced causes of anemia including defects of erythropoiesis, hemoglobin structure, RBC-enzymes and blood coagulation, hemolysis, immune disorders, infection, inflammation, extraintestinal hemorrhage, nephropathy were - according to the available data - unlikely to cause the anemia in our patients. Thus their anemia is thought to be due to age-inadequate cow's milk nutrition leading to 1) low intake, decreased absorption/bioavailability and increased intestinal loss of iron, and 2) GLB which induced blood loss following mechanical irritation of the gastric mucosa and vomiting causing high gastric pH and decrease in duodenal iron absorption. CONCLUSION: The anemia in our patients is due to both exaggerated feeding with cow's milk and adverse effects of GLBs. This hypothesis is supported by the finding that, after erythrocyte transfusion, iron substitution, age-adapted nutrition and GLB-dissolution, the anemia did not recur. We propose to include GLB in the differential diagnosis of anemia in cow's milk fed small children.


Subject(s)
Anemia, Iron-Deficiency/etiology , Bezoars/complications , Emigrants and Immigrants , Milk , Mucus , Anemia, Iron-Deficiency/diagnosis , Anemia, Iron-Deficiency/drug therapy , Animals , Austria , Bezoars/diagnosis , Combined Modality Therapy , Female , Ferrous Compounds/therapeutic use , Gastric Lavage , Humans , Infant , Stomach/diagnostic imaging , Ultrasonography
4.
J Neuroendocrinol ; 25(7): 617-25, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23387476

ABSTRACT

Delta-like 1 homologue (DLK1), also known as preadipocyte factor-1, fetal antigen 1 or pG2, is a transmembrane protein belonging to the epidermal growth factor-like superfamily. The protein becomes soluble and biologically active after cleavage of the tumour necrosis factor-α-converting enzyme. DLK1 is involved in the differentiation of several cell types, including adipocytes. Lack of the dlk1 gene in mice results in adiposity and a polymorphism within the gene encoding DLK1 has been associated with obesity. The dlk1 gene is expressed in restricted areas of the central nervous system with an enrichment of cell bodies expressing DLK1 mRNA in the hypothalamus. Goat and rabbit antisera to DLK1 were used to study the cellular localisation and chemical identity of DLK1-immunoreactive neuronal cell bodies in rat hypothalamus. DLK1 immunoreactivity was demonstrated in the cell bodies of the suprachiasmatic, supraoptic, paraventricular, arcuate nuclei and in the lateral hypothalamus. At the subcellular level, DLK1 immunoreactivity was observed in the cell soma and dendrites, although not in axonal fibres or nerve terminals. Double-labelling of sections from the lateral hypothalamic/perifornical area of colchicine-treated rats (a treatment that increases the content of immunoreactive material in the cell soma) showed that DLK1 was present in the virually all orexin- and dynorphin-containing neurones. By contrast, DLK1 was not demonstrated in any melanin-concentrating hormone or cocaine- and amphetamine-regulated transcript-containing neurones of the lateral hypothalamic/perifornical area. The presence of DLK1 in a population of lateral hypothalamic neurones suggests a functional role for DLK1 in orexin/hypocretin/dynorphin neurones.


Subject(s)
Hypothalamic Area, Lateral/metabolism , Intercellular Signaling Peptides and Proteins/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Membrane Proteins/metabolism , Nerve Tissue Proteins/metabolism , Neurons/metabolism , Neuropeptides/metabolism , Animals , Hypothalamic Area, Lateral/cytology , Immune Sera , Intercellular Signaling Peptides and Proteins/immunology , Male , Membrane Proteins/immunology , Orexins , Rats , Rats, Sprague-Dawley
5.
J Neuroendocrinol ; 24(12): 1492-504, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22775435

ABSTRACT

MicroRNAs (miRNAs) are short (∼22 nucleotides) noncoding RNA molecules that post-transcriptionally repress the expression of protein-coding genes by binding to 3'-untranslated regions of the target mRNAs. To identify miRNAs selectively expressed within the hypothalamus, a part of the brain that controls vital bodily functions, we employed locked nucleic acid (LNA)-fluorescent in situ hybridisation (FISH). The expression pattern of the mature miRNAs miR-7a, miR-7b, miR-137 and miR-153 in mouse brain tissue sections was investigated. Although all studied miRNAs were present in the hypothalamus, miR-7a, was the only miRNA found to be enriched in the hypothalamus, with low or no expression in other parts of the central nervous system (CNS). Within the hypothalamus, strong miR-7a expression was distinct and restricted to some hypothalamic nuclei and adjacent areas. miR-7a expression was particularly prominent in the subfornical organ, as well as the suprachiasmatic, paraventricular, periventricular, supraoptic, dorsomedial and arcuate nuclei. Identical expression patterns for miR-7a were seen in mouse and rat hypothalamus. By combining LNA-FISH with immunohistochemistry, it was shown that miR-7a was preferentially present in small orexigenic neuropeptide Y/agouti-related protein-containing-neurones located in the ventromedial aspect of the arcuate nucleus but not in large pro-opiomelanocortin/cocaine- and amphetamine-regulated transcript-containing anorexigenic neurones of the ventrolateral part of the arcuate nucleus. The limited and distinct expression of miR-7a in the CNS suggests that miR-7a has a role in post-transcriptional regulation in hypothalamic neurones. Particularly strong expression of miR-7a in neurones located in the ventromedial division of the arcuate nucleus, a subregion with a weak blood-brain barrier, raises the possibility that miR-7a is influenced by circulating hormones and is a regulator of the genes involved in body weight control.


Subject(s)
Gene Expression Regulation , Hypothalamus/metabolism , In Situ Hybridization, Fluorescence/methods , MicroRNAs/genetics , Oligonucleotides/pharmacology , Animals , Body Weight/genetics , Immunohistochemistry , Male , Mice , Mice, Inbred C57BL , MicroRNAs/metabolism , Rats , Rats, Sprague-Dawley , Tissue Distribution
6.
Neuroscience ; 171(1): 62-78, 2010 Nov 24.
Article in English | MEDLINE | ID: mdl-20736052

ABSTRACT

Purines such as ATP act as extracellular messengers through specific purinergic receptors. Three different classes of purinergic receptors have been identified and termed P1, P2X and P2Y. The purinergic receptor subunit P2X2 is a ligand-gated ion channel that is widely expressed by neurons in the central nervous system. The aim of this study was to study the cellular localization and to identify the chemical phenotypes of ionotropic P2X2 receptor (P2X2R)-containing neurons in the rat mediobasal hypothalamus by immunohistochemistry using three different P2X2R antisera, with special reference to neurons that influence food intake and body weight. P2X2R immunoreactivity was mainly observed in cell bodies and neural extensions located in the ventromedial part of the hypothalamic arcuate nucleus, a subregion of the nucleus with a weak blood-brain barrier (BBB). At the subcellular level, P2X2R immunoreactivity was located to the periphery of individual cells, likely representing the plasma membrane. Many P2X2R-immunoreactive cell bodies in the arcuate nucleus contained the orexigenic peptides neuropeptide Y (NPY) and agouti-related protein (AgRP), and the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD). In contrast, P2X2R immunoreactive cell bodies of the arcuate nucles only occasionally contained the anorexigenic peptides α-melanocyte-stimulating hormone (α-MSH) or cocaine- and amphetamine-regulated transcript (CART), or the opioid peptide dynorphin (DYN). There was no evidence for colocalization of P2X2R with somatostatin or neuronal nitric oxide synthase (nNOS) in neurons of the arcuate nucleus. In the parvocellular part of the paraventricular nucleus, P2X2R was demonstrated in some corticotropin-releasing hormone (CRH), thyrotropin-releasing hormone (TRH) and CART-containing neurons. In some cell bodies of the lateral hypothalamic area P2X2R was colocalized with DYN. The presence of P2X2R immunoreactivity in primarily orexigenic NPY/AgRP/GABA-containing neurons of the arcuate nucleus suggests that extracellular ATP has a regulatory action on this neuronal population located in a strategic position of the brain.


Subject(s)
Appetite Regulation/physiology , Hypothalamus/cytology , Neurons/physiology , Receptors, Purinergic P2X2/metabolism , Agouti-Related Protein/metabolism , Animals , Appetite Regulation/drug effects , Cell Count/methods , Cell Membrane/metabolism , Dynorphins/metabolism , Glutamate Decarboxylase/metabolism , Male , Nerve Tissue Proteins/metabolism , Neurons/cytology , Rats , Rats, Sprague-Dawley , Receptors, Purinergic P2X2/genetics , alpha-MSH/metabolism
7.
J Neuroendocrinol ; 22(2): 75-82, 2010 Feb.
Article in English | MEDLINE | ID: mdl-20025627

ABSTRACT

Nociceptin/orphanin FQ (N/OFQ), an endogenous peptide agonist of the opioid N/OFQ receptor, has been implicated in the regulation of energy balance. In the present study, we have used immunohistochemistry to investigate the cellular localisation and colocalisation of N/OFQ-immunoreactive cell bodies in hypothalamic regions containing neurones producing orexigenic or anorexigenic transmitters. In colchicine-treated rats, N/OFQ immunoreactivity was demonstrated in many cell bodies of the arcuate nucleus (Arc), paraventricular nucleus (PVN) and lateral hypothalamic area (LHA). Double-labelling revealed that N/OFQ was present in some neurones located in the ventrolateral part of the Arc producing pro-opiomelanocortin, as shown by the presence of the anorexigenic peptides alpha-melanocyte-stimulating hormone (alpha-MSH) and cocaine- and amphetamine-regulated transcript and, occasionally, in single neurones of the ventrolateral Arc producing orexigenic agouti-related peptide, but not neuropeptide Y. N/OFQ immunoreactivity was also demonstrated in a few tyrosine hydroxylase- or dynorphin (DYN)-containing neurones in the dorsomedial part of the Arc. In the parvocellular PVN, N/OFQ was demonstrated in some thyrotrophin-releasing hormone- or DYN-, but not corticotrophin-releasing hormone-containing neurones. Most N/OFQ-immunoreactive neurones in the LHA contained orexin- and DYN, but not melanin-concentrating hormone. The results obtained, demonstrating the presence of N/OFQ in some alpha-MSH- and in many orexin-containing neurones, suggest a functional relationship between these neuropeptides and N/OFQ in the control of feeding behaviour and body weight.


Subject(s)
Arcuate Nucleus of Hypothalamus/metabolism , Feeding Behavior/physiology , Hypothalamic Area, Lateral/metabolism , Neurons/metabolism , Opioid Peptides/metabolism , Paraventricular Hypothalamic Nucleus/metabolism , Animals , Hypothalamus/metabolism , Immunohistochemistry , Intracellular Signaling Peptides and Proteins/metabolism , Male , Neuropeptides/metabolism , Orexins , Rats , Rats, Sprague-Dawley , alpha-MSH/metabolism , Nociceptin
8.
Leukemia ; 23(7): 1264-9, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19212332

ABSTRACT

In the management of the childhood acute lymphoblastic leukemia (ALL), 5% of failures are due to induction death and treatment-related deaths in first complete remission. We retrospectively analyzed the incidence, pattern and causes of death and its risk factors for 896 children with ALL enrolled into five Austrian (A) Berlin-Frankfurt-Münster (BFM) trials between 1981 and 1999. The estimated 10-year cumulative incidence of death significantly decreased from 6+/-1% (n=16/268) in trials ALL-BFM-A 81 and ALL-A 84 to 2+/-1% (n=15/628) in trials ALL-BFM-A 86, 90 and 95 (P=0.006). A significant reduction of death was evident during induction therapy (2.2% in trials ALL-BFM-A 81 and ALL-A 84 and 0.2% in trials ALL-BFM-A 86, 90 and 95, P=0.001). Of 31 patients, 21 (68%) patients died from infectious and 10 (32%) from noninfectious complications. Treatment in trial ALL-BFM-A 81, infant age and female gender were independent predictors of an enhanced risk for death. Conclusively, we found a progressive reduction of death rates that may be explained by the increasing experience in specialized hemato-oncologic centers and improved supportive and intensive care. We also identified a distinct subset of patients who are especially prone to death and may need a special focus when receiving intense chemotherapy.


Subject(s)
Antineoplastic Agents/toxicity , Precursor Cell Lymphoblastic Leukemia-Lymphoma/mortality , Remission Induction , Cause of Death , Child , Female , Humans , Male , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy
10.
Mol Ecol Resour ; 9(2): 604-6, 2009 Mar.
Article in English | MEDLINE | ID: mdl-21564704

ABSTRACT

Six novel polymorphic microsatellite loci are presented for the grass snake (Natrix natrix), a species with declining populations in many regions. The number of alleles per locus ranged from two to seven. Four dice snake (Natrix tessellata) microsatellites were polymorphic in the grass snake with three to four alleles. At two loci, the expected heterozygosity differed significantly from observed heterozygosity. Cross-amplification of the grass snake markers in the dice snake showed two polymorphic microsatellites with two and four alleles.

11.
Neuroscience ; 154(3): 1121-31, 2008 Jun 26.
Article in English | MEDLINE | ID: mdl-18479830

ABSTRACT

Considerable evidence suggests that dynorphin participates in the regulation of energy balance. In this study, we have used immunohistochemistry to investigate in detail the cellular localization of pro-dynorphin (DYN) immunoreactive cell bodies in the mediobasal hypothalamus with special reference to neurons producing orexigenic or anorexigenic transmitters. In colchicine-treated rats, DYN immunoreactivity was demonstrated in many cell bodies of the arcuate nucleus (Arc). Double-labeling revealed that DYN immunoreactivity was present in approximately 30% of pro-opiomelanocortin (POMC) neurons in the ventrolateral Arc as shown by presence of alpha-melanocyte-stimulating hormone (alpha-MSH) and cocaine- and amphetamine-regulated transcript (CART). In contrast, DYN immunoreactivity was not demonstrated in agouti-related peptide (AgRP)- or neuropeptide Y (NPY) -containing neurons in the ventromedial aspect of the Arc. Dynorphin immunoreactivity was also colocalized with the vesicular acetylcholine (ACh) transporter (VAChT; a marker for cholinergic neurons) in the cell soma of Arc POMC neurons. Brainstem POMC neurons in the commissural part of the solitary tract nucleus (NTS) were devoid of DYN immunoreactivity, whereas DYN immunoreactivity was detected in a few NPY-containing NTS neurons and cholinergic DMX neurons. Our results showing presence of DYN together with alpha-MSH in a subpopulation of hypothalamic POMC neurons further point to the neurochemical heterogeneity of hypothalamic POMC neurons. The results suggest a role for DYN in control of energy balance by mediating the effect of peripheral hormones such as leptin and insulin.


Subject(s)
Arcuate Nucleus of Hypothalamus/metabolism , Dynorphins/metabolism , Neurons/metabolism , Pro-Opiomelanocortin/metabolism , Animals , Arcuate Nucleus of Hypothalamus/drug effects , Brain Stem/cytology , Brain Stem/drug effects , Brain Stem/metabolism , Cerebral Ventricles/drug effects , Cerebral Ventricles/metabolism , Colchicine/pharmacology , Energy Metabolism/drug effects , Energy Metabolism/physiology , Hypothalamus, Middle/drug effects , Hypothalamus, Middle/metabolism , Immunohistochemistry , Male , Median Eminence/drug effects , Median Eminence/metabolism , Nerve Tissue Proteins/metabolism , Neurons/drug effects , Rats , Rats, Sprague-Dawley , Ventromedial Hypothalamic Nucleus/drug effects , Ventromedial Hypothalamic Nucleus/metabolism , alpha-MSH/metabolism
12.
Cell Death Differ ; 14(3): 534-47, 2007 Mar.
Article in English | MEDLINE | ID: mdl-16888645

ABSTRACT

Protein kinase-B (PKB) and its target, the forkhead transcription factor like 1 (FKHRL1)/FoxO3a, have been suggested as regulators of neurotrophin-mediated cell survival in neuronal cells. We analyzed human neuroblastoma cells and found that FKHRL1 was phosphorylated, suggesting its inactivation. To study FKHRL1 function, we infected SH-EP and NB15 cells with a 4OH-tamoxifen-regulated FKHRL1(A3)ER(tm) transgene. Activation of FKHRL1 promoted cytochrome-c release and caspase-dependent apoptosis. FKHRL1 induced TRAIL and the BH3-only proteins Noxa and Bim, implicating both extrinsic and intrinsic death pathways. However, expression of dnFADD did not inhibit FKHRL1-induced cell death, whereas Bcl2 protected against apoptosis. This excluded the death-receptor pathway and suggested that cell death decision is regulated by Bcl2-rheostat. Importantly, RNAi knockdown of Noxa or Bim decreased apoptosis, indicating that Noxa and Bim cooperate to mediate FKHRL1-induced cell death. We conclude that Noxa and Bim establish a connection between FKHRL1 and mitochondria, and that both BH3-only proteins are critically involved in FKHRL1-induced apoptosis in neuroblastoma.


Subject(s)
Apoptosis Regulatory Proteins/metabolism , Forkhead Transcription Factors/metabolism , Gene Expression Regulation, Neoplastic , Membrane Proteins/metabolism , Mitochondria/metabolism , Neuroblastoma/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Proto-Oncogene Proteins/metabolism , 3-Phosphoinositide-Dependent Protein Kinases , Apoptosis , Apoptosis Regulatory Proteins/genetics , Bcl-2-Like Protein 11 , Caspases/metabolism , Cell Death , Fas-Associated Death Domain Protein/metabolism , Fas-Associated Death Domain Protein/physiology , Forkhead Box Protein O3 , Forkhead Transcription Factors/genetics , Humans , Membrane Proteins/genetics , Models, Biological , Peptide Fragments/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Protein Serine-Threonine Kinases/metabolism , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , Receptors, TNF-Related Apoptosis-Inducing Ligand/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , TNF-Related Apoptosis-Inducing Ligand/metabolism , Tamoxifen/analysis , Tamoxifen/pharmacology , Transduction, Genetic , fas Receptor/metabolism , fas Receptor/physiology
13.
J Chem Neuroanat ; 29(2): 93-111, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15652697

ABSTRACT

Activation of 5-HT1A receptors results in a variety of physiological responses, depending on their localization on neurons with different phenotypes in the brain. This study investigated the localization of 5-HT1A receptor mRNA and 5-HT1A receptor immunoreactivity in cell bodies of the rat septal complex using in situ hybridization and immunohistochemistry. In adjacent sections of the medial septum/diagonal band of Broca (MSDB), the distribution of cell bodies expressing 5-HT1A receptor mRNA was closely related to cells labeled with oligonucleotide probes to GAD (glutamic acid decarboxylase), VAChT (vesicular acetylcholine transporter) or parvalbumin mRNA. Using antiserum to GAD and antibodies to GABA, 5-HT1A receptor immunoreactivity was demonstrated in a majority of GABAergic cells in the MSDB. 5-HT1A receptor-immunoreactive GABAergic cells in the MSDB were also demonstrated to contain the calcium-binding protein parvalbumin, a marker for septohippocampal projecting GABAergic neurons. In the lateral septum, 5-HT1A receptor immunoreactivity was colocalized with the calcium-binding protein calbindin D-28k, a marker for septal GABAergic somatospiny neurons. 5-HT1A receptor immunoreactivity was also detected in a subpopulation of VAChT-containing cholinergic neurons of the MSDB. In MSDB neurons, colocalization of 5-HT1A and 5-HT2A receptor immunoreactivities was demonstrated. These observations suggest that serotonin via 5-HT1A receptors may represent an important modulator of hippocampal transmission important for cognitive and emotional functions through actions on both GABAergic and cholinergic neurons of the rat septal complex. In addition, 5-HT may exert its effects in the MSDB via cells expressing both 5-HT1A and 5-HT2A receptors.


Subject(s)
Acetylcholine/metabolism , Diagonal Band of Broca/metabolism , Neurons/metabolism , Receptor, Serotonin, 5-HT1A/genetics , Septal Nuclei/metabolism , gamma-Aminobutyric Acid/metabolism , Animals , Biomarkers , Calbindins , Diagonal Band of Broca/cytology , Efferent Pathways/physiology , Glutamate Decarboxylase/genetics , Glutamate Decarboxylase/metabolism , Hippocampus/physiology , Immunohistochemistry , Male , Membrane Transport Proteins/genetics , Neural Inhibition/physiology , Neurons/cytology , Parvalbumins/genetics , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT1A/metabolism , Receptor, Serotonin, 5-HT2A/metabolism , S100 Calcium Binding Protein G/metabolism , Septal Nuclei/cytology , Synaptic Transmission/physiology , Vesicular Acetylcholine Transport Proteins
14.
J Neuroendocrinol ; 16(7): 589-604, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15214862

ABSTRACT

gamma-Aminobutyric acid (GABA), the major inhibitory neurotransmitter in the brain, acts via two different type of GABA receptors. GABA(A) receptors are composed of five subunits that belong to eight different classes. Depending on their subunit composition, distinct pharmacological and electrophysiological properties are obtained. GABA is produced in certain hypothalamic neurones known to be involved in control of feeding behaviour. We report the detailed immunohistochemical localization of four GABA(A)R alpha subunits in hypothalamic regions associated with the regulation of feeding behaviour. Immunoreactive structures for all studied GABA(A)R alpha subunits were observed in the hypothalamus, but with subunit-specific staining patterns. GABA(A)R alpha(1) immunoreactivity was most prominent in the dorsomedial hypothalamic nucleus and in the lateral hypothalamic area (LHA), whereas GABA(A)R alpha(2), alpha(3) and alpha(5) subunits exhibited particularly strong immunoreactivity in the ventromedial hypothalamic nucleus. In comparison, GABA(A)R alpha subunit immunoreactivities were generally weak in the arcuate nucleus. In the ventromedial part of the arcuate nucleus, neuropeptide Y- and agouti-related peptide-containing cell bodies, which also are known to be GABAergic, were immunoreactive for only the GABA(A)R alpha(3) subunit, whereas pro-opiomelanocortin- and cocaine- and amphetamine-regulated transcript- containing cell bodies located in the ventrolateral subdivision of the arcuate nucleus, showed GABA(A)R alpha(1), alpha(2) and alpha(3) subunit immunoreactivity. In the LHA, GABA(A)R alpha(3) subunit immunoreactivity was demonstrated in both melanin-concentrating hormone (MCH) and orexin-containing neurones. In addition, MCH neurones contained GABA(A)R alpha(2) immunoreactivity. In neurones of the tuberomammillary nucleus, GABA(A)R alpha(2) and alpha(5) subunits were colocalized with histidine decarboxylase, a marker for histamine-containing neurones.


Subject(s)
Feeding Behavior/physiology , Hypothalamus/metabolism , Intracellular Signaling Peptides and Proteins , Receptors, GABA-A/metabolism , Animals , Arcuate Nucleus of Hypothalamus/cytology , Arcuate Nucleus of Hypothalamus/metabolism , Body Weight/physiology , Carrier Proteins/metabolism , Histidine Decarboxylase/metabolism , Hypothalamic Area, Lateral/cytology , Hypothalamic Area, Lateral/metabolism , Hypothalamic Hormones/metabolism , Hypothalamus/cytology , Immunohistochemistry , Intracellular Membranes/physiology , Male , Melanins/metabolism , Nerve Tissue Proteins/metabolism , Neurons/metabolism , Neuropeptides/metabolism , Orexin Receptors , Orexins , Paraventricular Hypothalamic Nucleus/cytology , Paraventricular Hypothalamic Nucleus/metabolism , Pituitary Hormones/metabolism , Pro-Opiomelanocortin/metabolism , Protein Subunits/classification , Protein Subunits/metabolism , Rats , Rats, Sprague-Dawley , Receptors, G-Protein-Coupled , Receptors, GABA-A/classification , Receptors, Neuropeptide , Tissue Distribution
15.
Brain Res Mol Brain Res ; 125(1-2): 129-39, 2004 Jun 18.
Article in English | MEDLINE | ID: mdl-15193430

ABSTRACT

A mutation in the mouse tub gene causes a phenotype characterized by maturity-onset obesity, blindness and deafness. The role of the intact tubby protein and the pathogenesis resulting in the phenotype of tub/tub mice remain largely unknown. In this study, we have investigated whether obese tub/tub mice exhibit altered expression levels for agouti-related protein (AGRP) or glutamic acid decarboxylase-65 (GAD65) in body weight-regulating neurons of the hypothalamic arcuate nucleus. In situ hybridization revealed that AGRP, but not GAD65 mRNA levels, were significantly lower in obese tub/tub mice as compared to tub/+ mice. The lower levels of AGRP mRNA in the arcuate nucleus of tub/tub mice were paralleled by lower fluorescence intensity and numbers of AGRP- and neuropeptide Y (NPY)-immunoreactive (ir) nerve fibers and terminals in the arcuate, ventromedial, dorsomedial hypothalamic nuclei and perifornical and lateral hypothalamic areas. No obvious differences in GAD65-ir nerve fibers and terminals could be detected. Measurements of daily food intake revealed that tub/tub mice displayed progressively higher food consumption as compared to lean tub/+ littermates over a 15-day observation period. When moved to an unfamiliar environment, e.g. a novel cage, daily food intake was initially lower in tub/tub mice than in tub/+ mice suggesting that tub/tub mice may be more sensitive to psychogenic stress. The results together show that tub/tub mice are hyperphagic and exhibit, within the hypothalamic arcuate nucleus, a depressed expression of neuropeptides involved in the regulation of feeding behavior.


Subject(s)
Arcuate Nucleus of Hypothalamus/metabolism , Down-Regulation , Obesity/metabolism , Proteins/metabolism , RNA, Messenger/metabolism , Adaptor Proteins, Signal Transducing , Agouti-Related Protein , Animals , Arcuate Nucleus of Hypothalamus/cytology , Eating , Female , Glutamate Decarboxylase/genetics , Glutamate Decarboxylase/metabolism , In Situ Hybridization , Intercellular Signaling Peptides and Proteins , Isoenzymes/genetics , Isoenzymes/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Proteins/genetics , Stress, Psychological
16.
Brain Res Mol Brain Res ; 120(1): 22-9, 2003 Dec 12.
Article in English | MEDLINE | ID: mdl-14667573

ABSTRACT

The family of G protein-coupled receptors (GPCRs) is one of the largest protein families in the mammalian genome. Receptors belonging to this class mediate the effects of very diverse ligands and are responsible for signaling events by affecting the activities of enzymes and ion channels. Here we describe the cloning and identification of GPCR-2037, a novel and previously not identified member of the large family of GPCRs. This orphan GPCR displays several typical features of family A type of GPCRs and shows highest homology with the galanin receptors 2 and 3. In rat brain, in situ hybridization showed that expression of GPCR-2037 mRNA was exclusively localized to neurons of the habenular complex. The expression was particularly prominent in the medial habenular nucleus, whereas the lateral habenular nucleus exhibited a lower number of labeled cells. The restricted and unique expression pattern of GPCR-2037 in the rat brain suggests a role for this orphan GPCR in the habenular complex, a brain structure implicated in the modulation of various physiological functions. Further studies involving the identification of the GPCR ligand will enable the functional characterization of this orphan receptor and its role in regulating the habenular complex.


Subject(s)
Habenula/cytology , In Situ Hybridization/methods , Neurons/metabolism , Receptors, G-Protein-Coupled/genetics , Amino Acid Sequence , Animals , Autoradiography , Base Sequence , Blotting, Northern , Brain/anatomy & histology , Brain/metabolism , Cloning, Molecular , Conserved Sequence , Humans , Male , Mice , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Receptors, G-Protein-Coupled/metabolism
18.
J Neuroendocrinol ; 15(1): 1-14, 2003 Jan.
Article in English | MEDLINE | ID: mdl-12535164

ABSTRACT

Gamma-aminobutyric acid (GABA) interacts with hypothalamic neuronal pathways regulating feeding behaviour. GABA has been reported to stimulate feeding via both ionotropic GABA(A) and metabotropic GABA(B) receptors. The functional form of the GABA(B) receptor is a heterodimer consisting of GABA(B) receptor-1 (GABA(B)R1) and GABA(B) receptor-2 (GABA(B)R2) proteins. Within the heterodimer, the GABA-binding site is localized to GABA(B)R1. In the present study, we used an antiserum to the GABA(B)R1 protein in order to investigate the cellular localization of GABA(B)R1-immunoreactive neurones in discrete hypothalamic regions implicated in the control of body weight. The colocalization of GABA(B)R1 immunoreactivity with different chemical messengers that regulate food intake was analysed. GABA(B)R1-immunoreactive cell bodies were found in the periventricular, paraventricular (PVN), supraoptic, arcuate, ventromedial hypothalamic, dorsomedial hypothalamic, tuberomammillary nuclei and lateral hypothalamic area (LHA). Direct double-labelling showed that glutamic acid decarboxylase (GAD)-positive terminals were in close contact with GABA(B)R1-containing cell bodies located in all these regions. In the ventromedial part of the arcuate nucleus, GABA(B)R1-immunoreactive cell bodies were found to contain neuropeptide Y, agouti-related peptide (AGRP) and GAD. In the ventrolateral part of the arcuate nucleus, GABA(B)R1-immunoreactive cell bodies were shown to contain pro-opiomelanocortin and cocaine- and amphetamine-regulated transcript. In the LHA, GABA(B)R1 immunoreactivity was present in both melanin-concentrating hormone- and orexin-containing cell populations. In the tuberomammillary nucleus, GABA(B)R1-immunoreactive cell bodies expressed histidine decarboxylase, a marker for histamine-containing neurones. In addition, GAD and AGRP were found to be colocalized in some nerve terminals surrounding GABA(B)R1-immunoreactive cell bodies in the parvocellular part of the PVN. The results may provide a morphological basis for the understanding of how GABA regulates the hypothalamic control of food intake and body weight via GABA(B) receptors.


Subject(s)
Feeding Behavior/physiology , Hypothalamus/chemistry , Neurons/chemistry , Receptors, GABA-B/analysis , Animals , Antibodies , Arcuate Nucleus of Hypothalamus/chemistry , Arcuate Nucleus of Hypothalamus/physiology , Body Weight/physiology , Dorsomedial Hypothalamic Nucleus/chemistry , Dorsomedial Hypothalamic Nucleus/physiology , Hypothalamic Area, Lateral/chemistry , Hypothalamic Area, Lateral/physiology , Hypothalamus/physiology , Hypothalamus, Anterior/chemistry , Hypothalamus, Anterior/physiology , Male , Paraventricular Hypothalamic Nucleus/chemistry , Paraventricular Hypothalamic Nucleus/physiology , Rats , Rats, Sprague-Dawley , Receptors, GABA-B/immunology , Ventromedial Hypothalamic Nucleus/chemistry , Ventromedial Hypothalamic Nucleus/physiology , gamma-Aminobutyric Acid/physiology
19.
Article in English | MEDLINE | ID: mdl-14720189

ABSTRACT

The quantification of slow-growing mycobacteria such as Mycobacterium tuberculosis or M. bovis from in vitro and in vivo samples is complicated by their long generation time, their ability to form aggregates, and their capacity to persist in a state of dormancy. We compared different methods for the establishment of growth curves for broth cultures of M. bovis bacille Calmette-Guérin (BCG). A quantitative TaqMan-PCR yielded results comparable with those obtained by protein quantification and measurement of the ATP content of the cultures. The quantitative TaqMan-PCR furthermore turned out to be particularly suitable for the measurement of multiplication of BCG within eukaryotic cells. Furthermore, it is a fast method allowing an estimation of the mycobacterial load in tissue long before colony counts can be obtained.


Subject(s)
Mycobacterium bovis/physiology , Mycobacterium tuberculosis/physiology , Polymerase Chain Reaction/veterinary , Animals , Cells, Cultured/physiology , Culture Media , DNA, Bacterial/analysis , Female , Guinea Pigs , Macrophages/microbiology , Male , Mycobacterium bovis/genetics , Mycobacterium tuberculosis/genetics , Predictive Value of Tests , Stem Cells
20.
Br J Haematol ; 115(3): 545-53, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11736934

ABSTRACT

Childhood acute lymphoblastic leukaemia (ALL) is a malignancy with the potential to infiltrate the liver, spleen, lymph nodes and brain. Such extramedullary presentation is important for understanding the biology of childhood ALL and also for developing new prognostic parameters. A potential mechanism in the trafficking of leukaemia cells is the interaction of the chemokine receptor CXCR4, which is expressed on ALL cells, and its ligand stromal cell-derived factor-1 (SDF-1), produced by stromal cells in bone marrow and extramedullary organs. Functionality of CXCR4 was demonstrated by a high correlation between cell surface density of CXCR4 and transendothelial migration of leukaemia blasts towards a gradient of SDF-1 (r = 0.73, P = 0.001). Inhibition of SDF-1-induced migration by an anti-CXCR4 monoclonal antibody (78.33 +/- 23.86% inhibition) evidenced the specificity of CXCR4 to SDF-1. In order to evaluate clinical significance of CXCR4 expression, lymphoblasts from the bone marrow of 73 patients with and without extramedullary organ infiltration were compared. Multiparameter flow cytometry revealed that lymphoblasts from patients with high extramedullary organ infiltration, defined as ultrasonographically measured enlargement of liver or spleen, expressed the CXCR4 receptor at higher fluorescence intensity (median 66.12 +/- 66.17) than patients without extramedullary organ infiltration (median 17.56 +/- 19.29; P < 0.001). Consequently, high expression of CXCR4 was strongly predictive for extramedullary organ involvement, independently of the peripheral lymphoblast count. Highest CXCR4 expression was seen in mature B ALL (median 102.74 +/- 92.13; P < 0.003), a disease characterized by a high incidence of extramedullary bulky disease. As high expression of the chemokine receptor CXCR4 predicts extramedullary organ infiltration in childhood ALL, we suggest that CXCR4 and its ligand play an essential role in extramedullary invasion.


Subject(s)
Leukemic Infiltration/diagnosis , Lymphocytes/metabolism , Precursor Cell Lymphoblastic Leukemia-Lymphoma/metabolism , Receptors, CXCR4/analysis , Antibodies, Monoclonal/pharmacology , Biomarkers/analysis , Chemokine CXCL12 , Chemokines, CXC/metabolism , Chemotaxis, Leukocyte/drug effects , Child , Child, Preschool , Female , Flow Cytometry/methods , Humans , Immunohistochemistry/methods , Infant , Jurkat Cells , Liver/pathology , Male , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Prognosis , Receptors, CXCR4/immunology , Receptors, CXCR4/metabolism , Spleen/pathology
SELECTION OF CITATIONS
SEARCH DETAIL
...