scholarly journals Role of nerve growth factor in the development of rat sympathetic neurons in vitro. I. Survival, growth, and differentiation of catecholamine production.

1977 ◽  
Vol 75 (3) ◽  
pp. 694-704 ◽  
Author(s):  
L L Chun ◽  
P H Patterson
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neuronal Survival ◽  
Sympathetic Neurons ◽  
Cell Types ◽  
Neonatal Rat ◽  
Nerve Growth ◽  
Dose Dependent Fashion ◽  
Lipid Phosphate

To study the effect of nerve growth factor (NGF) on neuronal survival, growth, and differentiation, cultures of dissociated neonatal rat sympathetic neurons virtually free of other cell types were maintained for 3-4 wk. In the absence of NGF, the neurons did not survive for more than a day. Increased levels of NGF increased neuronal survival and growth (total protein and total lipid phosphate); saturation occurred at 0.5 microgram/ml 7S NGF. Neuronal differentiation examined by measuring catecholamine (CA) production from tyrosine also depended on the level of NGF in the culture medium. As the NGF concentration was raised, CA production per neuron, per nanogram protein, or per picomole lipid phosphate increased until saturation was achieved between 1 and 5 microgram/ml 7S NGF. Thus, NGF induces neuronal survival, growth, and differentiation of CA production in a dose-dependent fashion. Neuronal growth and differentiation were quantitatively compared in the presence of the high and low molecular weight forms of NGF; no significant functional differences were found.

scholarly journals Role of nerve growth factor in the development of rat sympathetic neurons in vitro. III. Effect on acetylcholine production.

1977 ◽  
Vol 75 (3) ◽  
pp. 712-718 ◽  
Author(s):  
L L Chun ◽  
P H Patterson
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Rat Heart ◽  
Neuronal Survival ◽  
Sympathetic Neurons ◽  
Cell Monolayer ◽  
Heart Cell ◽  
Heart Cells ◽  
Nerve Growth ◽  
Rat Heart Cells

The effect of nerve growth factor (NGF) on the development of cholinergic sympathetic neurons was studied in cultures grown either on monolayers of dissociated rat heart cells or in medium conditioned by them. In the presence of rat heart cells the absolute requirement of neurons for exogenous NGF was partially spared. The ability of heart cells to support neuronal survival was due at least in part to production of a diffusable NGF-like substance into the medium. Although some neurons survived on the heart cell monolayer without added NGF, increased levels of exogenous NGF increased neuronal survival until saturation was achieved at 0.5 microgram/ml 7S NGF. The ability of neurons to produce acetylcholine (ACh) from choline was also dependent on the level of exogenous NGF. In mixed neuron-heart cell cultures, NGF increased both ACh and catecholamine (CA) production per neuron to the same extent; saturation occurred at 1 microgram/ml 7S NGF. As cholinergic neurons developed in culture, they became less dependent on NGF for survival and ACh production, but even in older cultures approximately 40% of the neurons died when NGF was withdrawn. Thus, NGF is as necessary for survival, growth, and differentiation of sympathetic neurons when the neurons express cholinergic functions as when the neurons express adrenergic functions (4, 5).

scholarly journals Pit formation and rapid changes in surface morphology of sympathetic neurons in response to nerve growth factor.

1981 ◽  
Vol 90 (1) ◽  
pp. 176-180 ◽  
Author(s):  
J L Connolly ◽  
S A Green ◽  
L A Greene
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Surface ◽  
Sympathetic Neurons ◽  
Cell Types ◽  
Coated Pits ◽  
Nerve Growth ◽  
Pheochromocytoma Cells ◽  
Pit Formation ◽  
Transmission Electron

Scanning and transmission electron microscope studies were carried out on the rapid cell surface responses of cultured newborn rat sympathetic neurons to nerve growth factor (NGF), a substance that promotes their survival and differentiation. The somas of sympathetic neurons continuously exposed to NGF or deprived of the factor for 4-5 h have a very smooth surface. After readdition of NGF to the latter type of cultures, there is rapidly initiated a transient, sequential change in the cell surface. Microvilli and small ruffles appear within 30 s and are most prominent by 1 min. By 3 min of exposure, the microvilli and ruffles decrease in prominence, and by 7 min the somal surface is again smooth. By 30 s after NGF readdition, as increase in the number of 60- tp 130-nm coated pits is also detectable. This increase reaches a maximum of about threefold from 0.5 to 3 min and then gradually decreases. Alterations in the surface did not occur on the nonneuronal cell types present in the cultures and were not observed in response to another basic protein (cytochrome c) or to physical manipulation. Changes in cell surface architecture induced by NGF in normal sympathetic neurons and, as previously described, in PC12 pheochromocytoma cells indicate that such responses may present or reflect primary events in the mechanism of the factor's action.

scholarly journals Role of nerve growth factor in the development of rat sympathetic neurons in vitro. II. Developmental studies.

1977 ◽  
Vol 75 (3) ◽  
pp. 705-711 ◽  
Author(s):  
L L Chun ◽  
P H Patterson
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Time Course ◽  
Sympathetic Neurons ◽  
Protein A ◽  
Cell Number ◽  
Similar Time ◽  
Nerve Growth ◽  
Virtual Absence

Adrenergic sympathetic neurons were grown for 4 wk in submaximal and saturating concentrations of nerve growth factor (NGF) in the virtual absence of non-neuronal cells. In 0.2 or 5 microgram/ml 7S NGF, the neurons gradually decreased in number during the first week, although fewer neurons died at the higher level. No significant change in cell number was observed thereafter. Total neuronal protein, a measure of cell growth, increased linearly with age in both concentrations of NGF. At each age, neurons in high NGF exhibited greater growth per cell than those in low NGF. The ability of neurons to produce catecholamine (CA) increased dramatically during the second and third weeks in both concentrations of NGF, and along a similar time-course, although neurons in submaximal NGF developed a lesser capacity for CA production. As neurons developed in culture, they became less dependent on NGF for survival and CA production, but even in older cultures, approximately 50% of the neurons died when NGF was withdrawn.

scholarly journals Temporal analysis of events associated with programmed cell death (apoptosis) of sympathetic neurons deprived of nerve growth factor.

1993 ◽  
Vol 123 (5) ◽  
pp. 1207-1222 ◽  
Author(s):  
T L Deckwerth ◽  
E M Johnson
Keyword(s):  
Cell Death ◽  
Protein Synthesis ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Programmed Cell Death ◽  
Dna Fragmentation ◽  
Molecular Mechanisms ◽  
Sympathetic Neurons ◽  
Neonatal Rat ◽  
Nerve Growth

The time course of molecular events that accompany degeneration and death after nerve growth factor (NGF) deprivation and neuroprotection by NGF and other agents was examined in cultures of NGF-dependent neonatal rat sympathetic neurons and compared to death by apoptosis. Within 12 h after onset of NGF deprivation, glucose uptake, protein synthesis, and RNA synthesis fell precipitously followed by a moderate decrease of mitochondrial function. The molecular mechanisms underlying the NGF deprivation-induced decrease of protein synthesis and neuronal death were compared and found to be different, demonstrating that this decrease of protein synthesis is insufficient to cause death subsequently. After these early changes and during the onset of neuronal atrophy, inhibition of protein synthesis ceased to halt neuronal degeneration while readdition of NGF or a cAMP analogue remained neuroprotective for 6 h. This suggests a model in which a putative killer protein reaches lethal levels several hours before the neurons cease to respond to readdition of NGF with survival and become committed to die. Preceding loss of viability by 5 h and concurrent with commitment to die, the neuronal DNA fragmented into oligonucleosomes. The temporal and pharmacological characteristics of DNA fragmentation is consistent with DNA fragmentation being part of the mechanism that commits the neuron to die. The antimitotic and neurotoxin cytosine arabinoside induced DNA fragmentation in the presence of NGF, supporting previous evidence that it mimicked NGF deprivation-induced death closely. Thus trophic factor deprivation-induced death occurs by apoptosis and is an example of programmed cell death.

scholarly journals Characterization of apoptosis in cultured rat sympathetic neurons after nerve growth factor withdrawal

1994 ◽  
Vol 124 (4) ◽  
pp. 537-546 ◽  
Author(s):  
SN Edwards ◽  
AM Tolkovsky
Keyword(s):  
Cell Death ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Sympathetic Neurons ◽  
Chromatin Condensation ◽  
Nerve Growth ◽  
Culture Age ◽  
Membrane Breakdown

Sympathetic neurons depend on nerve growth factor (NGF) for their survival both in vivo and in vitro. In culture, the neurons die after NGF withdrawal by an autonomous cell death program but whether these neurons die by apoptosis is under debate. Using vital DNA stains and in situ nick translation, we show here that extensive chromatin condensation and DNA fragmentation occur before plasma membrane breakdown during the death of NGF-deprived rat sympathetic neurons in culture. Furthermore, kinetic analysis of chromatin condensation events within the cell population is consistent with a model which postulates that after NGF deprivation nearly all of the neurons die in this manner. Although the dying neurons display membrane blebbing, cell fragmentation into apoptotic bodies does not occur. Apoptotic events proceed rapidly at around the time neurons become committed to die, regardless of neuronal culture age. However the duration of NGF deprivation required to commit neurons to die, and the rate at which apoptosis occurs, increase with culture age. Thus, within the first week of culture, apoptosis is the predominant form of cell death in sympathetic neurons.

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