bluuebunny/crossref_metadata_2025_split huggingface dataset:
Localization of constitutive and hyperthermia‐inducible heat shock mRNAS (hsc70 and hsp70) in the rabbit cerebellum and brainstem by non‐radioactive in situ hybridization
Authors: Foster J. A. , Rush S. J. , Brown I. R. 
Published: January 2005

AbstractNeural expression of constitutive hsc70 mRNA and hyperthermia‐inducible hsp70 mRNA is examined using radioactive and non‐radioactive in situ hybridization procedures. A strong induction of hsp70 mRNA was noted in cell populations in cerebellar layers and in the brainstem which demonstrated expression of mRNA encoding proteolipid protein, an oligodendrocyte marker. The non‐radioactive in situ hybridization procedure using digoxigenin (DIG)‐UTP‐labeled riboprobes permitted improved signal localization, and stress‐inducible hsp70 mRNA was detected at the cytoplasmic cap areas of individual oligodendrocytes. Cell types which express constitutive members of the hsc/hsp70 multigene family were also identified. Neurons in the brainstem and in the deep white matter and molecular layer of the cerebellum showed expression of hsc70 mRNA while signal was not detected in adjacent glial cells. A neuron‐specific enolase riboprobe aided in the identification of neuronal cell types. The non‐radioactive DIG riboprobe revealed that hsc70 mRNA was highly localized to the cyto‐plasm of individual neurons. High constitutive levels of hsc70 in certain neurons may dampen hsp70 induction after hyperthermia in these cell populations. © 1995 Wiley‐Liss, Inc.
Exposure of tumor necrosis factor‐α to luminal membrane of bovine brain capillary endothelial cells cocultured with astrocytes induces a delayed increase of permeability and cytoplasmic stress fiber formation of actin
Authors: Deli M. A. , Descamps L. , Dehouck M.‐P. , Cecchelli R. , Joó F. , Ábrahám C. S. , Torpier G. 
Published: January 2005

AbstractTumor necrosis factor‐α (TNF‐α), a proinflammatory cytokine, has long been known to be involved in the pathogenesis of central nervous system infections and of certain neurodegenerative diseases. However, the possible role of the blood‐brain barrier (BBB), the active interface between the blood circulation and brain tissue, remained unknown during these pathological conditions. In our in vitro reconstructed BBB model, 1‐hr exposure of recombinant human TNF‐α (in concentrations of 50, 250, and 500 U/ml, respectively) to the luminal membrane of bovine brain capillary endothelial cells (BBCEC) did not change significantly the transendothelia: flux of either sucrose (m.w. 342 Da), or inulin (m.w. 5 kDa) up to 4 hr (early phase), except for a slight decrease (P< 0.05) in sucrose permeation at 2–4 hr with the highest dose of TNF‐α On the other hand, at 16 hr after the 1‐hr challenge with TNF‐α (delayed phase) at all 3 concentrations, significant increase was induced in the permeability of BBCEC monolayers for both markers. These changes of permeability were accompanied by a selective reorganization of F‐actin filaments into stress fibers, while the intracellular distribution of vimentin remained similar to the control. These results suggest that BBCEC can respond directly to TNF‐α by a delayed increase of permeability and reorganization of actin filaments. ©1995 Wiley‐Liss, Inc.
Purine and pyrimidine nucleotides activate distinct signalling pathways in PC12 cells
Authors: De Souza L. R. , Moore H. , Raha S. , Reed J. K. 
Published: January 2005

AbstractThe role of extracellular nucleotides in intracellular signalling and neurosecretion was assessed in PC12 cells. Activation of phospholipase C and increased [Ca2+]i were mediated by purinoceptors with an agonist potency profile, ATP ∼ UTP > 2‐methylthioadenosine triphosphate (2‐MeSATP), typical of P2U. ATP also evoked a rapid acidification followed by a more gradual alkalinization (measured with 2′,7′‐biscarboxyethyl‐5(6)‐carboxyfluorescein (BCECF)), while UTP induced only a gradual alkalinization. The amiloride analogue 5‐(N‐ethyl‐N‐isopropyl) amiloride (EIPA) attenuated the alkalinization phase suggesting activation of the Na+/H+ exchanger by ATP and UTP. Using bisoxonol and [3H]tetraphenylphosphonium ([3H]TPP+) as potential‐sensitive probes, we showed that while ATP rapidly depolarized PC 12 cells in an Na+ ‐dependent manner, UTP evoked a much reduced and delayed response. The potency profile (ATP ∼ 2‐MeSATP ∼ adenosine 5′‐0‐(3thiotriphosphate) (ATPγS) ≫ UTP, α, beta;‐methyleneATP) suggested involvement of a receptor subtype distinct from P2U. Secretion of endogenous dopamine was also assessed. Those nucleotides that induced depolarization (ATP, 2‐MeSATP, ATPγS) were also the most potent secretagogues. UTP was ineffective. Our results suggest that ATP stimulates distinct purinoceptor subtypes and induces neurosecretion through the activation of multiple signalling path ways. © 1995 Wily‐Liss, Inc.
On identifying a second molecular antagonistic mechanism operative at the glycine receptor
Authors: Galvez‐Ruano E. , Lipkowitz K. B. , Aprison M. H. 
Published: January 2005

AbstractWe used molecular modeling techniques to examine six reported antagonists of glycine with varying K; values against strychnine. We found the data suggest two groups operating with different mechanisms. In group 1 (strychnine, brucine, Pitrazepin, and bicuculline methobromide) the antagonist contains two or three sites that can electrostatically bind to the three comparable groups of opposite charge in the recognition site where the natural neurotransmitter binds, thus opening the chloride channel. In addition, when in this position, the antagonist is able to also block the now opened chloride channel with a different portion of its structure. In many cases, this involves an interaction between a carbonyl group, on the antagonist and the guanidinium group of arginine which is part of the polypeptide segment of the outer mouth of the chloride channel (Grenningloh et al., Nature 330:2526, 1987). In group 2 (R5135 and 1,5‐diphenyl‐3,7‐diazaadamantan‐9‐ol) the antagonist contains charged sites but when one of these molecules attaches to the recognition site, the chloride channel is not opened. In addition, R5135 contains a carbonyl group which attaches to arginine as pointed out in the text, whereas 1,5‐diphenyl‐3,7‐diazaadamantan‐9‐ol contains a phenyl group that can block the channel. © 1995 Wiley‐Liss, Inc
Neuronal differentiation of P19 embryonal carcinoma cells in defined media
Authors: Yao M. , Bain G. , Gottlieb D. I. 
Published: January 2005

AbstractThe P19 embryonal carcinoma cell line is a useful model system for analyzing the factors that regulate neuronal differentiation. In order to analyze the extrinsic factors that are involved in differentiation, it is necessary to carry out experiments in fully defined media. Here we have investigated the neuronal differentiation of P19 cells in two defined media. Cells that are propagated and induced with retinoic acid in standard serum‐containing medium are capable of differentiating into neuron‐like cells in N2 medium. Dividing fibroblast‐like cells also appeared in these cultures. After about 10 days in culture in N2 medium, the great majority of neuron‐like cells died. On the other hand, culturing induced cells in N2 medium for 5 days and then switching to a defined medium consisting of Neurobasal medium plus B27 supplement allowed the neuron‐like cells to survive for prolonged periods of time. This defined medium thus provides a suitable system for analyzing extrinsic factors that affect the survival and differentiation of P19 neurons. P19 cells induced with retinoic acid and plated in N2 were exposed to bFGF and EGF, which are known to be mitogens for neuronal precursor cells. Both growth factors were mitogenic for a subpopulation of the induced cells. In separate experiments, cells cultured in N2 in the presence of RA were induced to differentiate into neuron‐like cells. © 1995 Wiley‐Liss, Inc.
GM1 ganglioside rescues substantia nigra pars compacta neurons and increases dopamine synthesis in residual nigrostriatal dopaminergic neurons in MPTP‐treated mice
Authors: Schneider J. S. , Kean A. , DiStefano L. 
Published: January 2005

AbstractGM1 ganglioside has been shown to stimulate recovery of the damaged dopamine system under a number of different circumstances. In addition to rescue of damaged dopamine neurons, the present study assessed the ability of GM1 to enhance the synthesis of dopamine in remaining nigrostriatal neurons following 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP) exposure. There was a significantly greater accumulation of L‐dopa 30 min after aromatic amino acid decarboxylase inhibition with NSD‐1015 (100 mg/Kg) and an increase in the ratio of L‐dopa to dopamine in MPTP + GM1‐treated mice than in mice that received only MPTP. This effect of GM1 on dopamine synthesis was dependent upon the degree of initial damage to the nigrostriatal dopamine system. That is, the GM1 effect on dopamine synthesis could not be demonstrated in mice with greater than 95% striatal dopamine loss and 75% substantia nigra dopamine neuron loss. These results suggest that in addition to previously reported effects of GM1 on rescue and repair of dopaminergic neurons, GM1 may also have the ability to enhance dopamine synthesis in residual dopaminergic neurons. Direct effects on dopamine neurochemistry may contribute to functional improvements seen after GM1 treatment in various models of parkinsonism. © 1995 Wiley‐Liss, Inc.
Differentially expressed genes after peripheral nerve injury
Authors: Gillen C. , Gleichmann M. , Spreyer P. , Müller H. W. 
Published: January 2005

AbstractIn an attempt to identify genes associated with Wallerian degeneration and peripheral nerve regeneration we have performed differential hybridization screening of a cDNA library from crushed rat sciatic nerve (7 days postlesion) using radioactively labeled cDNA prepared from poly(A)+ RNA of normal vs. crushed nerve. Screening of 5,000 randomly selected colonies yielded 24 distinct clones that were regulated following nerve injury. Fifteen of the differentially expressed sequences could be classified as induced, whereas 9 sequences appeared to be repressed at 1 week postcrush. Sequencing and computer‐assisted sequence comparison revealed 3 classes of regulated cDNA clones representing (1) novel gene sequences (8 clones) including 3 transcripts containing a repetitive “brain identifier” (ID) element; (2) identified genes (7 clones) with previously undetected expression in the peripheral nervous system (PNS), such as apolipoprotein D, peripheral myelin protein 22kD (PMP22), SPARC (secreted protein, acidic and rich in cysteine), sulfated glycoprotein SGP‐1, apoferritin, decorin, and X16/SRp20; and (3) identified genes (9 clones) with known expression in the PNS including, e.g., the myelin protein P0, γ‐actin, vimentin, a‐tubulin, chargerin II, and cytochrome c‐oxidase subunit I. Northern blot and polymerase chain reaction analyses with RNA from crushed and transected nerve demonstrated that sequences with related function, like the group of myelin genes, cytoskeleton genes, genes involved in RNA processing and translation, in lipid transport or energy metabolism showed closely related temporal patterns of expression during nerve degeneration and regeneration. Finally, we compared the differentially expressed genes identified at 7 days after crush injury (this investigation) with the regulated sequences isolated previously by De Leon et al. (J Neurosci Res 29:437‐488, 1991) from a 3 day postcrush sciatic nerve cDNA library. © 1995 Wiley‐Liss, Inc.
Gene therapy in the central nervous system: Direct versus indirect gene delivery
Authors: Ridet J.‐L. , Privat A. 
Published: January 2005

AbstractOver the last decade, the combination of molecular biology and cell transplantation techniques has given rise to a powerful method for gene therapy. The implantation of genetically modified cultured cells has been extensively used in the central nervous system (CNS) in various experimental models of neurologic disorders. More recently, viral and chemical methods have been developed to further efforts to shuttle transgenes into the relatively inaccessible brain. Adenoviral and liposomal synthetic vectors carry transgenes into neural tissue in situ and are beginning to show promise as new methods for CNS therapy. © 1995 Wiley‐Liss, Inc.
Dystroglycan expression in the wild type and mdx mouse neural retina: Synaptic colocalization with dystrophin, dystrophin‐related protein but not laminin
Authors: Montanaro F. , Carbonetto S. , Campbell K. P. , Lindenbaum M. 
Published: January 2005

AbstractAlpha‐ and β‐dystroglycan (α‐ and β‐DG) are members of a dystrophin‐associaced glycoprotein complex (DGC) in skeletal muscle which binds to agrin and laminin, and has been postulated to be involved in myoneural synapse formation. The absence of functional dystrophin in Duchenne muscular dystrophy (DMD) and in one of its animal models, the mdx mouse, leads to a reduction of α‐ and β‐DG in muscle, and is often associated with mental retardation and abnormal retinal synaptic transmission in DMD. Using immunohistochemistry, we find that a‐ and β‐DG are expressed in the outer plexiform layer of both wild type and mdx retina, where both dystrophin and dystrophin‐related protein (DRP), but not laminin are present. In situ hybridization identifies two neuronal populations, photoreceptors and retinal ganglion cells, that express DG mRNA. α‐ and β‐DG are also expressed in the inner limiting membrane and around blood vessels where they colocalize with laminin and DRP. Western blot analysis revealed the expression of several dystrophin isoforms in wild type and mdx retina, possibly explaining the unaltered expression of α‐ and β‐dystroglycan in the mdx central nervous system (CNS). Our results support the hypothesis that a‐ and β‐DG can interact with dystrophin and DRP in the CNS and perform functions analogous to those of the DGC in muscle. © 1995 Wiley‐Liss, Inc.
Developmental expression of calmodulin mRNA and protein in regions of the postnatal rat brain
Authors: Berry F. , Brown I. R. 
Published: January 2005

AbstractThe expression of calmodulin (CaM) protein and mRNA was analyzed in specific regions of the rat brain during postnatal development. CaM levels in the adult brain were more abundant in the cerebral hemispheres and thalamus compared to brain stem and superior plus inferior colliculus. All brain regions contained higher CaM protein and mRNA levels than in non‐neural tissues such as the kidney. During postnatal development of the brain, maximal levels of CaM protein and CaM I mRNAs were attained at day 10 or 15. Protein levels declined thereafter in the adult in all regions except the thalamus. With respect to products of the rat CaM I gene, the 4.0 kb neural transcript demonstrated a pronounced increase during postnatal development, whereas the 1.8 kb message showed little change. © 1995 Wiley‐Liss, Inc.
Trophic and protective effects of growth/differentiation factor 5, a member of the transforming growth factor‐β superfamily, on midbrain dopaminergic neurons
Authors: Krieglstein K. , Suter‐Crazzolara C. , Hötten G. , Pohl J. , Unsicker K. 
Published: January 2005

AbstractGrowth/differentiation factor 5 (GDF5) is a novel member of the transforming growth factor‐β (TGF‐β) superfamily of multifunctional cytokines. We show here that GDF5 is expressed in the developing CNS including the mesencephalon and acts as a neurotrophic, survival promoting molecule for rat dopaminergic midbrain neurons, which degenerate in Parkinson's disease. Recombinant human GDF5 supports dopaminergic neurons, dissected at embryonic day (E) 14 and cultured for 8 days under serum‐free conditions, to almost the same extent as TGF‐β, and is as effective as glial cell line‐derived neurotrophic factor (GDNF), two established trophic factors for midbrain dopaminergic neurons. In contrast to TGF‐β and GDNF, GDF5 augments numbers of astroglial cells in the cultures, suggesting that it may act indirectly and through pathways different from those triggered by TGF‐β and GDNF. GDF5 also protects dopaminergic neurons against the toxicity of N‐methylpyridinium ion (MPP+), which selectively damages dopaminergic neurons through mechanisms currently debated in the etiology of Parkinson's disease (PD). GDF5 may therefore now be tested in animal models of PD and might become useful in the treatment of PD. © 1995 Wiley‐Liss, Inc.
Convergent influence of the central nucleus of the amygdala and the paraventricular hypothalamic nucleus upon brainstem autonomic neurons as revealed by c‐fos expression and anatomical tracing
Authors: Petrov T. , Krukoff T. L. , Jhamandas J. H. 
Published: January 2005

AbstractCombinations of anatomical tracing with detection of Fos (the protein product of the immediate early gene c‐fos) consequent to the stimulation of the central nucleus of the amygdala were used to explore the possibility that the hypothalamic paraventricular nucleus participates in the activation of brainstem neurons in the nucleus of the solitary tract and ventrolateral medulla. After injections of the anterograde tracer Phaseolus vulgaris leucoagglutinin in the paraventricular nucleus, labeled fibers and varicosities were found to impinge on catecholaminergic and non‐catecholaminergic Fos‐positive neurons in the brainstem. After injections of a retrograde tracer in the nucleus of the solitary tract or ventrolateral medulla, we observed that some of the Fos‐positive neurons within the parvocellular paraventricular nucleus that project to the brainstem were catecholaminergic or oxytocinergic.The results indicate that direct and indirect inputs from the amygdala may influence the activity of autonomic neurons in the brainstem. The paraventricular nucleus, via its direct projections onto catecholaminergic and non‐catecholaminergic neurons, may participate in activation of brainstem neurons. Activated catecholaminergic and oxytocinergic parvocellular neurons in the paraventricular nucleus may be involved in the transmission of autonomic signals from the amygdala toward the brainstem. ©1995 Wiley‐Liss, Inc.
CD9 of mouse brain is implicated in neurite outgrowth and cell migration in vitro and is associated with the α6/β1 integrin and the neural adhesion molecule L1
Authors: Schmidt C. , Künemund V. , Wintergerst E. S. , Schmitz B. , Schachner M. 
Published: January 2005

AbstractWe describe here a novel monoclonal antibody (mab INTRODUCTION H6) which recognizes CD9, an integral cell surface constituent previously described in cells of the hematopoietic lineage and involved in the aggregation of platelets. Mab H6 was raised against membranes of immature mouse astrocytes and reacted with a protein of 25–27 kD in detergent extracts of adult mouse brain membranes. Sequence analysis of the N‐terminal amino acids revealed an identity of 96% with CD9 from mouse kidney. CD9 was localized in the central and peripheral mouse nervous systems: in the spinal cord of 11‐day‐old mouse embryos, CD9 was strongly expressed in the floor and roof plates. In the adult mouse sciatic nerve, myelin sheaths were highly CD9immunoreactive. Mab H6 reacted with the cell surfaces of both glial cells and neurons in culture and inhibited migration of neuronal cell bodies, neurite fasciculation and outgrowth of astrocytic processes from cerebellar microexplants. Neurite outgrowth from isolated small cerebellar neurons was increased in the presence of mab H6 on substrate‐coated laminin, but not on substrate‐coated poly‐L‐lysine. Addition of mab H6 elicited an increase in intracellular Ca2+ concentration in these cells on substrate‐coated laminin. Immunoprecipitates of CD9 from cultured mouse neuroblastoma N2A cells contained the α6/β1 integrin. Moreover, preparations of CD9 immunoaffinity‐purified from adult mouse brain using a mab H6 column contained the neural adhesion molecule L1, but not other neural adhesion molecules. CD9 bound to L1, but not to NCAM or MAG. Both the α6/β1 integrin and L1 could be induced to coredistribute with CD9 on the surface of cultured neuroblastoma N2A cells. The combined observations suggest that CD9 can associate with L1 and α6/β1 integrin to influence neural cell interactions in vitro. © 1996 Wiley‐Liss, Inc.
Impaired synaptic functions with aging as characterized by decreased calcium influx and acetylcholine release
Authors: Tanaka Y. , Hasegawa A. , Ando S. 
Published: January 2005

AbstractAge‐related alterations of presynaptic functions were studied in terms of acetylcholine (ACh) synthesis and release using synaptosomes isolated from mouse brain cortices. The following three findings were obtained: (1) Choline acetyltransferase activity and ACh production rate remained constant throughout all ages tested. This observation, obtained with synaptosomes, was not consistent with data reported for brain slices (Gibson GE, Peterson C: J Neurochem 37:978–984, 1981). Various conditions, such as low glucose or membrane depolarization, modulated ACh synthesis to similar extents in young and aged synaptosomes. (2) Depolarization‐induced release of ACh from synaptosomes significanty decreased in the senescent stage. The fraction of ACh released from aged synaptosomes was less than that released from young synaptosomes, although the ACh contents in the synaptosomes did not change with age. (3) Calcium influx induced by depolarization was lower in the synaptosomal preparations from aged mice than in those from young mice. A strong positive correlation was observed between the amounts of ACh released and the increased calcium levels when the data for all preparations, both from young and aged mice, were plotted. This indicates that diminished calcium influx may cause the reduced ACh release by aged synapses. The present study provides evidence for an age‐related decrease in presynaptic functions, that is, a reduction in calcium influx via voltage‐dependent calcium channels followed by a decreased ACh release from synapses despite an abundance of ACh within the synapses. © 1996 Wiley‐Liss, Inc.
Tics in a patient with Parkinson's disease
Authors: Shale Heidi , Fahn Stanley , Mayeux Richard 
Published: January 2005

AbstractA patient with Gilles de la Tourette syndrome later developed Parkinson's disease in middle age. This was accompanied by a marked reduction in the frequency of tics but levodopa toxicity exacerbated the tics. The dopamine hypothesis of tic disorders is supported by this observation.
Electrophoresis and immunoblot of cerebrospinal fluid proteins in spasmodic torticollis
Authors: Cullis Paul A. , Townsend Laurace , Lewitt Peter , Pomara Nunzio , Reitz Darlene 
Published: January 2005

AbstractProtein patterns of cerebrospinal fluid (CSF) from patients with spasmodic torticollis (ST) were investigated to determine whether abnormalities previously reported could be detected and further identified. CSF was collected from 12 patients with ST and 6 normal controls. The CSF proteins were analyzed using sodium dodecyl sulfate (SDS) polyacrylmide gel electrophoresis and silver staining. In 11 of the 12 patients with ST, a CSF protein pattern was observed which differed from that in the controls. The identity of the abnormal proteins was ascertained by blotting and immunostaining with specific antisera to IgG and ceruloplasmin (Cp). CSF from 2 of 12 patients had distinct bands staining for IgG and 7 had abnormal immunostaining for Cp.