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B Meister

Publications and source records attributed to B Meister.

At least 55 records · Page 3Linked to original sources
PubMed1998

Estrogen down-regulates mRNA encoding the exocytotic protein SNAP-25 in the rat pituitary gland.

Exocytosis is dependent on specific proteins that are located at the secretory granule membrane, in the cytoplasm or at the plasma membrane. The mRNA expression of synaptosomal-associated protein of 25 kDa (SNAP-25) isoforms SNAP-25a and SNAP-25b, vesicle associated membrane protein (VAMP) 2, mammalian homologue of unc-18 (munc-18) and Hrs-2 was studied in the pituitary of ovariectomized rats after subcutaneous insertion of capsules containing estrogen or placebo using in situ hybridization. Estrogen treatment (0.25 mg estradiol) significantly decreased SNAP-25a (32%; 10%) and SNAP-25b (25%; 22%) mRNA levels in the anterior and intermediate lobes, respectively, whereas VAMP-2, munc-18 and Hrs-2 mRNA levels remained unchanged. The results suggest that estrogen selectively regulates SNAP-25 transcription in the pituitary gland, but leaves VAMP-2, munc-18 and Hrs-2 mRNA levels unaffected.

Adenosine Triphosphatases↗
PubMed1998

VAMP-1 and VAMP-2 gene expression in rat spinal motoneurones: differential regulation after neuronal injury.

Vesicle-associated membrane protein (VAMP; synaptobrevin) is involved in the molecular regulation of transmitter release at the presynaptic plasma membrane. VAMP exists in two isoforms, VAMP-1 and VAMP-2, which are transcribed from two separate genes and differentially expressed in the nervous system. In situ hybridization was used to examine whether VAMP isoform mRNA expression may be altered by experimental manipulations. The effect of nerve injury on VAMP-1 and VAMP-2 mRNA levels in motoneurones of the rat lumbar spinal cord was compared with lesion-induced changes in the expression of choline acetyl transferase (ChAT) and alpha-calcitonin gene-related peptide (alpha-CGRP) mRNA. After unilateral sciatic nerve transection (axotomy), VAMP-1 mRNA expression decreased significantly in parallel with a downregulation of ChAT mRNA in axotomized motoneurones compared with the corresponding motoneurones on the contralateral unlesioned side. There was a rapid decrease in VAMP-1 and ChAT mRNA levels at 2 days after axotomy, and at 7 days there was a 65% decrease in VAMP-1 mRNA and a 48% decrease in ChAT mRNA. VAMP-1 mRNA levels continued to decrease at 14 and 21 days, while ChAT mRNA levels had returned to normal at this time. In contrast, VAMP-2 and alpha-CGRP mRNA levels were upregulated in axotomized motoneurones. A significant increase for both VAMP-2 and alpha-CGRP mRNA levels was present 2 days after axotomy, and a maximum was reached after 7 days for alpha-CGRP mRNA (163%) and after 14 days for VAMP-2 mRNA (587%). Immunohistochemical analysis did not reveal any detectable changes in VAMP-1- or VAMP-2-like immunoreactivity in the motoneurone cell soma after axotomy. In the proximal end of the transected sciatic nerve, there was an increase in VAMP-1- and VAMP-2-LI, which was most prominent at 2 days after lesion. The results show that, in axotomized spinal motoneurones, VAMP-1 mRNA is downregulated and VAMP-2 mRNA is upregulated, indicating differential regulation of the two separate VAMP genes and differential roles for the two VAMP isoforms in the regulation of exocytosis after nerve injury.

Acetylcholine↗
PubMed1998

Decreasing reticulocyte counts associated with declining post-dose erythropoietin plasma levels in anaemia of prematurity.

UNLABELLED: A prospective sequential, multicentre trial was conducted to determine the association between erythropoietin (EPO) plasma levels and the erythropoietic response to recombinant human erythropoietin (r-HuEPO) during long-term treatment of premature infants. Twenty-nine infants, gestational ages 26-34 weeks and postnatal ages more than 14 days, received 600 IU r-HuEPO per kg per week divided into three doses subcutaneously for haemoglobin levels less than 120 g/l or haematocrit less than 36% over a period of 4 weeks. Eight additional patients were studied for a total of 10 weeks. EPO plasma concentrations and haematologic parameters were measured prior to the onset of treatment and at 2-weekly intervals thereafter. Treatment with r-HuEPO resulted in a median increase in corrected reticulocyte counts of 2.5% (range 0.2-4.6%) above patient's baseline, thereafter a decrease was observed. In the 8 patients followed for 10 weeks reticulocyte counts declined significantly during weeks 6-10 when compared with the first 4 weeks (p < 0.005). Median 72-hour post-dose EPO plasma levels increased significantly (p < 0.0001) to 57.3 mU/ml (range 5.0-160) above patient's baseline after the first injection, but declined progressively thereafter until they approached baseline values at week 10. CONCLUSION: R-HuEPO treatment after the first month was associated with a decrease in post-injection plasma levels and a decrease in erythropoietic response. This decrease in erythropoietin's efficacy and the decline observed in post-dose EPO plasma levels may be causally related.

Anemia↗
PubMed1998

Transcription factor STAT3 in leptin target neurons of the rat hypothalamus.

Leptin is an adipose tissue-derived hormone that regulates body weight via interactions with hypothalamic neuronal circuitries expressing specific leptin receptors (Ob-R). The Ob-Rs act via the JAK-STAT (Janus kinase-signal transducers and activators of transcription) pathway of signal transduction. Recent evidence suggests that primarily the transcription factor STAT3 mediates leptin's action in the hypothalamus. We have investigated the presence and cellular localization of STAT3 protein in the rat hypothalamus by means of indirect immunofluorescence histochemistry using a rabbit polyclonal STAT3 antiserum. The antiserum identified a 92-kDa protein using Western blotting on rat hypothalamic homogenates, corresponding to the expected size of STAT3. STAT3 immunoreactivity was demonstrated in Ob-R-containing neurons of the paraventricular nucleus (parvocellular part), periventricular nucleus, arcuate nucleus and in the lateral hypothalamic area. Direct double-labeling showed presence of STAT3 immunoreactivity in neuropeptide Y (NPY)-containing neurons of the ventromedial part of the arcuate nucleus and in proopiomelanocortin (POMC)-containing neurons of the ventrolateral part of the arcuate nucleus. The results provide an anatomical basis for a leptin action mediated by STAT3 in Ob-R-containing NPY and POMC neurons of the arcuate nucleus, as well as by Ob-R-containing neurons of the parvocellular paraventricular nucleus and lateral hypothalamic area.

Animals↗
PubMed1998

Ex vivo expansion of hematopoietic progenitor cells for clinical use.

The development of efficient stem and progenitor cell selection methods in combination with the development of hematopoietic growth factors facilitated the development of ex vivo expansion techniques. Currently, this novel domain of cellular therapy aims to generate stem and progenitor cells, as well as more differentiated post-progenitor cells and antigen-presenting dendritic cells. The feasibility of generating and transplanting hematopoietic progenitor cells ex vivo (using various cytokine combinations) has been successfully shown preclinically as well as clinically. Furthermore, cytokines (eg, Flt-3-ligand; thrombopoietin) have been identified that play important roles with regard to amplification of undifferentiated early hematopoietic cells. The use of lineage-specific cytokines such as granulocyte colony-stimulating factor and thrombopoietin facilitated the generation of large numbers of myeloid and megakaryocytic post-progenitor cells. The clinical usefulness of such ex vivo generated cells, however, has not yet been convincingly shown. Last, ex vivo expansion techniques can be used to generate large numbers of antigen-presenting dendritic cells from CD34+ peripheral blood progenitor cells that might be ideally used for immunotherapeutic approaches.

Cell Count↗
PubMed1997

Botulinum neurotoxin F, a VAMP-specific endopeptidase, inhibits Ca(2+)-stimulated GH secretion from rat pituitary cells.

Botulinum neurotoxin F (BoNTx F) is a zinc-dependent endopeptidase that causes proteolytic cleavage of the vesicle protein VAMP (vesicle-associated membrane protein). VAMP is an important component of the molecular machinery regulating docking and fusion of secretory vesicles with the target membrane. We have investigated presence of VAMP protein in cultured rat anterior pituitary cells. Confocal laser microscopy revealed presence of VAMP-like immunoreactivity in secretory granules of GH-containing cultured rat anterior pituitary cells. Using BoNTx F, we have investigated whether VAMP is involved in growth hormone (GH) secretion. Treatment of streptolysin-O permeabilized GH-secreting cells with BoNTx F (2.0 and 20 nM) significantly inhibited Ca(2+)-induced GH release. The results show that the secretory granules of rat anterior pituitary cell contain VAMP protein and suggest that VAMP is of importance in regulating Ca(2+)-mediated GH secretion.

Animals↗
PubMed1997

Vesicular acetylcholine transporter (VAChT) protein: a novel and unique marker for cholinergic neurons in the central and peripheral nervous systems.

Acetylcholine (ACh) is synthesized in nerve terminals from choline and acetyl coenzyme A by the cytoplasmic enzyme choline acetyltransferase (ChAT). The neurotransmitter is thereafter transported into synaptic vesicles, where it is stored until release. cDNA clones encoding a vesicular ACh transporter (VAChT) were recently isolated. In this paper, we report on the generation of highly specific goat polyclonal antisera to the rat VAChT protein by using a synthetic carboxy-terminal 20-amino-acid peptide sequence as an immunogen. Characterization of the antisera revealed recognition of VAChT, but not vesicular monoamine transporter (VMAT) protein, in transfected CV-1 cells. VAChT immunoreactivity was also detected in cells that endogenously express the protein, such as in PC12 cells and in primary cultures of spinal motoneurons. Absorption controls showed that the VAChT antisera could be completely blocked at the 10(-5) M concentration by cognate peptide used for immunization. The antisera cross-reacted with the VAChT protein in rat and mouse but not in guinea pig, rabbit, or cat. Immunohistochemistry and confocal laser microscopy, using the goat VAChT antisera, showed strong immunoreactivity in discrete fibers and neuronal cell bodies of the central and peripheral nervous systems. Within cell bodies and axonal nerve terminals, as well as in dendrites, the staining appeared granular, presumably representing labeling of synaptic vesicles containing ACh. In the rat central nervous system, VAChT-positive cell bodies were demonstrated in the cerebral cortex, striatum, septum, nucleus basalis, medial habenula, mesopontine complex, cranial, and autonomic and spinal motor nuclei and in the intermediomedial region near the central canal. High densities of VAChT-immunoreactive axonal fibers were encountered in areas such as the olfactory bulb, cerebral cortex, striatum, basal forebrain, amygdala, thalamus, hypothalamus including median eminence, hippocampal formation, superior colliculus, interpeduncular nucleus, and pedunculopontine and laterodorsal tegmental nuclei. In cranial and spinal motor nuclei, particularly large varicosities were seen in close proximity to the motoneuron cell somata and their proximal dendrites. In the peripheral nervous system, VAChT immunoreactivity was also detected in motor endplates of skeletal muscle as well as in fibers of sympathetic and parasympathetic abdominal ganglia, heart atrium, respiratory tract, gastrointestinal tract, pancreas, adrenal medulla, male genitourinary tract, and salivary and lacrimal glands. Direct double labeling revealed colocalization of VAChT and ChAT immunoreactivity in neurons. The results show that VAChT antisera represent novel and unique tools for the study of cholinergic neurons in the central and peripheral nervous systems.

Animals↗
PubMed1997

The effect of recombinant human erythropoietin on circulating hematopoietic progenitor cells in anemic premature infants.

In vitro and animal studies suggest that high concentrations of recombinant human erythropoietin (rHuEPO) might divert multipotent progenitors into erythroid maturation at the expense of granulocyte production. We determined whether changes of number and lineage commitment of peripheral blood progenitor cells occur in premature infants during therapy with rHuEPO. Thirty preterm infants were randomly assigned either to receive 300 IU of eopoetin alpha s.c. per kilogram body weight three times a week for four weeks or to a control group. At study entry and after two weeks of treatment the numbers of circulating BFU-E, granulocyte-macrophage colony-forming units (CFU-GM) and granulocyte-erythrocyte-macrophage-megakaryocyte CFU (CFU-GEMM) were analyzed by semisolid culture technique, CD34+ cells and early myeloid CD34+CD45RA- progenitors by flow cytometry. As compared with the control group, rHuEPO treatment did not exert any significant modulatory effect on numbers of CFU-GM, nor was there a significant change in numbers of BFU-E, CFU-GEMM, total-CFU, percentage of CD34+ or CD34+CD45RA- cells. Mean neutrophil count was not significantly reduced at any period during the study. Compared with the control group, the infants receiving rHuEPO had higher hematocrit values (p = 0.003) and absolute reticulocyte counts (p < 0.001). The median cumulative volume of blood transfused per kilogram per day was 0.86 ml (first quartile 0.5 ml; third quartile 1.1 ml) in the control group and 0 ml (first quartile 0 ml; third quartile 0.47 ml) in the rHuEPO group (p = 0.038). We conclude using a relatively high dose of rHuEPO in premature infants, no significant in vivo effect on circulating peripheral blood progenitor or neutrophil count could be detected.

Anemia↗
PubMed1997

Exocytotic proteins in enterochromaffin-like (ECL) cells of the rat stomach.

Proteins participating in vesicular docking and fusion have been identified in the nervous system. Such proteins appear to be important for the molecular regulation of exocytosis also in non-neuronal cells. The enterochromaffin-like (ECL) cells of the gastric acid-secreting (oxyntic) mucosa secrete histamine and chromogranin A-derived peptides, such as pancreastatin. Using immunohistochemistry, we have examined whether the ECL cells of the rat stomach, identified with antibodies to histidine decarboxylase (HDC, the histamine-forming enzyme), express the same exocytotic proteins as neurons. The ECL cells displayed immunoreactivity for synaptophysin, synaptotagmin III, vesicle-associated membrane protein-2 (VAMP-2), cysteine string protein (CSP), vesicular monoamine transporter-2 (VMAT-2), synaptosomal-associated protein of 25 kDa (SNAP-25), syntaxin, and Munc-18, but not for synaptotagmin I/II and VAMP-1. Synaptophysin and VMAT-2 could be detected not only in the ECL cells, but also in a population of HDC-negative cells. The demonstration of synaptotagmin III in only a limited number of ECL cells suggests the existence of a subpopulation of ECL cells. The results show that several exocytotic proteins, previously identified in neurons, are present in rat stomach ECL cells. Hence, proteins engaged in vesicular docking and in the fusion of granule/vesicle membrane with plasma membrane seem to exist in both neurons and endocrine cells.

Animals↗
PubMed1996

Effect of oxytocin on growth hormone release in vitro.

The effect of oxytocin (OT) on growth hormone (GH) secretion was investigated using dispersed rat anterior pituitary cells. OT dose-dependently inhibited GH secretion as well as GHRH-stimulated GH release. The inhibitory actions of OT on GH release were totally abolished by pretreatment with the OT-antagonist VAP 259. The peptides galanin and cholecystokinin did not affect the OT-induced inhibition on basal or GHRH-stimulated GH release. Several possible mechanisms by which OT may influence GH release are discussed.

Animals↗
PubMed1996

Expression of leptin receptor mRNA in the hypothalamic arcuate nucleus--relationship with NPY neurones.

The obese phentotype of ob/ob mice is linked to a mutation in the ob gene that results in expression of a truncated inactive protein. The ob gene product, leptin, is synthesized in adipose tissue and is a circulating factor that regulates body weight. Leptin receptors were recently cloned and a mutation in the leptin receptor gene in obese db/db mice was identified. Leptin receptor mRNA has been detected in brain, including hypothalamus, but the cellular localization has so far not been clarified. Here we report on the cellular localization of leptin receptor mRNA in the mouse brain using in situ hybridization. Strong hybridization was observed in the choroid plexus and hypothalamic arcuate nucleus. Weaker hybridization was detected in the hippocampal formation and in the cerebral cortex. Within the arcuate nucleus, cell bodies expressing leptin receptor mRNA were distributed in its ventromedial subdivision. Hybridization of semiadjacent sections with a probe to neuropeptide Y (NPY) showed a co-distribution of labelled cell bodies, suggesting the presence of leptin receptors on NPY-containing neurones.

Animals↗
PubMed1996

[The hormone leptin reduces body weight. A mutant gene makes the mouse obese].

The existence of mice (ob/ob mice) with a genetic defect causing obesity and type II diabetes has been known since 1950. The mutated ob gene was recently identified and characterized. The gene encodes a 167 amino acid protein that has been given the name leptin, from the greek word leptos, meaning thin. The functionally active hormone, which is synthesised and secreted by adipocytes, is lacking in homozygote ob/ob mice, causing an increase in body fat. Injection of recombinant leptin in ob/ob mice induces loss of fat due to decreased appetite and increased energy expenditure. The ob gene product leptin acts via binding sites in the hypothalamus, where the centre for appetite and satiety is located. Research is now focused on the identification, characterization and cloning of leptin-receptors. Other mice, also with a genetic defect causing obesity and type II diabetes, do not respond to leptin treatment and are therefore suspected to have defective leptin receptors.

Animals↗
PubMed1996

Changes in enkephalin and neuropeptide Y-like immunoreactivity in rabbit chromaffin tissues during perinatal development.

Enkephalin-like immunoreactivity (ENK-LI), neuropeptide Y (NPY)-LI and dopamine-beta-hydroxylase (DBH)-LI were found within the chromaffin cells of both the paraaortic body and the adrenal medulla of the newborn rabbit using immunohistochemistry. Cells positive to DBH-LI were abundant in both the paraaortic body and the adrenal medulla. ENK-LI positive cells were frequent in the paraaortic body, but more sparse in the adrenal medulla. A few cells staining for NPY-LI could be detected in both organs. Some nerve fibers within these organs also contained substance P-LI and calcitonin-gene related peptide-LI. The tissue contents of ENK-LI and NPY-LI, as measured by radioimmunoassay, increased after birth in the adrenal glands and were significantly higher than the fetal levels from 1 week of age. In the paraaortic body the lowest content of ENK-LI was found around birth, whereas the content of NPY-LI was highest at that time. With advancing postnatal age, the content of ENK-LI increased, whereas the content of NPY-LI decreased. At each age, there was a higher content of ENK-LI as compared to NPY-LI in both organs. This indicates that the synthesis of ENK-LI and NPY-LI in the paraaortic body is differently regulated during perinatal development.

Adrenal Medulla↗
PubMed1996

Differential subcellular localization of SNAP-25a and SNAP-25b RNA transcripts in spinal motoneurons and plasticity in expression after nerve injury.

Synaptosomal-associated protein of 25 kDa (SNAP-25) is involved in the molecular regulation of neurotransmitter release. SNAP-25 exists in two isoforms, which arise from alternative splicing of exon 5. In situ hybridization was used to examine whether SNAP-25 isoform mRNA expression may be altered by experimental manipulations. The effect of unilateral nerve injury on SNAP-25 mRNA levels was studied in motoneurons of the rat lumbar spinal cord. In all animals, SNAP-25a RNA transcripts were demonstrated in the nucleus of motoneurons, whereas SNAP-25b mRNA was present mainly in the cytoplasm. Cloning of the rat Snap gene intron spacing the alternative exon 5a and 5b sequences and generation of an intron-specific oligonucleotide probe used for in situ hybridization did not point to the presence of unspliced variants of SNAP-25b mRNA. After unilateral sciatic nerve transection (axotomy), SNAP-25a and SNAP-25b expression decreased in axotomized motoneurons compared with corresponding motoneurons on the unlesioned side. A significant decrease was demonstrated 2 days after axotomy, which reached a maximum after 7 days (62% for SNAP-25a and 67% for SNAP-25b), while levels had slightly recovered by 14 and 28 days. Ventral root avulsion also induced a decrease in levels of SNAP-25 RNA transcripts, suggesting that the axonal injury in itself was responsible for the down-regulation of Snap gene expression. This study shows that, in spinal motoneurons, SNAP-25a and SNAP-25b RNA transcripts have different subcellular localization and that levels of SNAP-25 RNA transcripts are down-regulated after axonal injury.

Animals↗
PubMed1996

Molecular components of the exocytotic machinery in the rat pituitary gland.

Several protein components that are essential for exocytotic membrane fusion in neurons have recently been identified. The expression and cellular localization of such protein components were examined in the rat pituitary gland. In situ hybridization using isoform-specific oligonucleotide probes to different exocytotic protein messenger RNAs (mRNAs) showed strong hybridization signal for synaptotagmin I, cysteine string protein (CSP), VAMP-2 (vesicle-associated membrane protein), cellubrevin, munc-18 (mammalian homologue of unc-18), SNAP-25a (synaptosomal-associated protein of 25 kDa), syntaxin 1A, syntaxin 4, syntaxin 5, and alpha-SNAP (soluble NSF attachment protein) in the anterior and intermediate, but not in the posterior lobe of the pituitary. Moderate to weak hybridization signal was detected for synaptotagmin III, SNAP-25b, and syntaxin 2 mRNA in the anterior and intermediate, but not in the posterior lobe of the pituitary. Synaptotagmin II, VAMP-1, syntaxin 1B, or syntaxin 3 mRNA expression could not be detected in any part of the pituitary gland. Immunofluorescence histochemistry in combination with confocal laser microscopy revealed that synaptotagmin-, VAMP-, CSP-, NSF-, and alpha-SNAP-like immunoreactivities (-LI) were present in granules of cells in the anterior and intermediate lobe, whereas SNAP-25- and syntaxin-LI were primarily located to the plasma membrane. Synaptotagmin-, VAMP-, CSP-, NSF-, alpha-SNAP-, SNAP-25- and syntaxin-LI were all present in nerve fibers of the posterior lobe. Within cells of the anterior lobe, colocalization could be demonstrated for synaptotagmin I/II- and synaptotagmin III-LI with ACTH-, GH-, PRL- and TSH-, but not FSH- or LH-LI, whereas VAMP, CSP-, NSF-, alpha-SNAP-, SNAP-25- and syntaxin-LI were demonstrated in all hormone-containing cell types of the anterior pituitary. The results show the presence of several protein components and their isoform-specific mRNAs in the rat pituitary gland, suggesting that these proteins, similar to their roles in regulation of synaptic neurotransmitter release, may participate in exocytotic events in endocrine pituitary cells and in neurosecretory nerve endings of the neurohypophysis.

Animals↗
PubMed1996

Combinatorial expression patterns of the connexins 26, 32, and 43 during development, homeostasis, and regeneration of rat teeth.

Gap junctions permit the exchange of regulatory molecules between cells and play important roles during organogenesis. The expression pattern of the gap junction proteins connexin 26, 32, and 43 was studied by immunohistochemistry in the developing, adult, and injured rat teeth. Connexins 32 and 43, but not the connexin 26, were detected during the late stages of embryonic tooth development (bell stage). Expression of connexin 32 was predominant in epithelial cells, whereas connexin 43 was more widely distributed and found in both epithelial and mesenchymal cells. During cytodifferentiation (early postnatal stages), both connexin 32 and 43 were expressed in the epithelial-derived ameloblasts, synthesizing and secreting the enamel matrix proteins. In mesenchyme, connexin 32 was observed only in differentiating odontoblasts, while connexin 43 was expressed in both differentiating and functional odontoblasts, which secrete the dentin matrix. In adult rat teeth, connexin 26 and 43 were expressed in the odontoblastic layer at low and high levels, respectively, while connexin 32 was absent from odontoblasts. Electron microscopy showed that connexin 43 was distributed exclusively at sites of contacts between odontoblasts. However, double immunostaining combined with confocal microscopy suggested an occasional overlap between odontoblasts and calcitonin gene-related peptide-positive nerve fibers. Denervation experiments showed that the expression of connexins in dental pulp was independent of innervation, whereas in injured teeth connexin 43 was upregulated in pulpal fibroblasts. Finally, cultured dental epithelial cells expressed both connexin 32 and 43, and connexin 43 was detected in cultured pulp fibroblasts in vitro, thus mimicking the in vivo distribution pattern of connexins. These results demonstrate that connexins are involved in tooth development and suggest that a given connexin may have distinct roles during odontogenesis and tooth homeostasis.

Animals↗