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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Neurochemical research 13 (1988), S. 611-616 
    ISSN: 1573-6903
    Keywords: Anti-oxidants ; catalase ; cultures ; free radicals ; neuronotrophic ; neurons ; pyruvate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The search for neuronotrophic factors addressing CNS neurons requires CNS neuronal cell cultures to quantitate putative effects on neuronal survival. Investigation of neurons dissociated from several embryonic CNS tissues have shown that their short-term survival requires supplementation of the culture medium with either pyruvate or the enzyme catalase. Pyruvate can be replaced with α-ketoglutarate or oxaloacetate, or with amino acids capable to transaminate to these three metabolites in the presence of exogenous α-ketoacid acceptors. Experiments were designed to evaluate the ability of cultured CNS neurons to utilized glucose as their primary source. We show that: (1) catalase requires the availability of glucose in the medium in order to exert its neuronal maintenance effect, (2) in the absence of catalase, the cells are unable to metabolize glucose through the tricarboxylic acid cycle, (3) catalase restores the neuronal ability to utilize glucose for oxydative metabolism, and renders redundant the use of other sources such as glutamate conversion to α-ketoglutarate, (4) graded concentrations of glucose in the medium affect in parallel these metabolic activities and the viability of the cultured neurons, and (5) anti-oxidant agents other than catalase mimic the catalase effects. We conclude that dissociated embryonic CNS neurons suffer from a block in glucose utilization which results from an imbalance between free radical attack and cellular defenses to it and speculate on a more general involvement of peroxidation damage in the trophic requirements for neuronal survival.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-6903
    Keywords: Ciliary neurotrophic factor ; mutagenesis ; secondeary protein structure ; biological activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Ciliary neurotrophic factor (CNTF) is a polypeptide that promotes the survival and/or differentiation of a number of neural cell types. Here we present a structural and functional analysis of the human CNTF molecule. Variant proteins were synthesized byEscherichia coli trnsformmed with mutant cDNA constructs, and purified by SDS-polyacrylamide gel electrophoresis and reverse phase high pressure liquid chromatography. Most variant CNTF proteins lacked neurotrophic activity, but two N-and C-terminal deletions (Δ2–14 and Δ173–200, respectively) actually displayed a several-fold increase in specific activity. Loss of biological activity was accompanied by changes in the alphahelical nature of CNTF as measured by circular dichroism. These data strengthen the proposed similarity between CNTF and the family of hematopoietic cytokines.
    Type of Medium: Electronic Resource
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