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on NGF for survival and CA production, but even in older cultures, approximately 50% of the neurons died when NGF was withdrawn.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 Nerve growth factor-induced neurite outgrowth in PC12 cells involves the coordinate induction of microtubule assembly and assembly-promoting factors.

1985 ◽  
Vol 101 (5) ◽  
pp. 1799-1807 ◽  
Author(s):  
D G Drubin ◽  
S C Feinstein ◽  
E M Shooter ◽  
M W Kirschner
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neurite Outgrowth ◽  
Pc12 Cells ◽  
Time Course ◽  
Microtubule Assembly ◽  
Nerve Growth ◽  
Peripheral Neurons ◽  
Associated Proteins

Nerve growth factor (NGF) regulates the microtubule-dependent extension and maintenance of axons by some peripheral neurons. We show here that one effect of NGF is to promote microtubule assembly during neurite outgrowth in PC12 cells. Though NGF causes an increase in total tubulin levels, the formation of neurites and the assembly of microtubules follow a time course completely distinct from that of the tubulin induction. The increases in microtubule mass and neurite extension closely parallel 10- and 20-fold inductions of tau and MAP1, proteins shown previously to promote microtubule assembly in vitro. When NGF is removed from PC12 cells, neurites disappear, microtubule mass decreases, and both microtubule-associated proteins return to undifferentiated levels. These data suggest that the induction of tau and MAP1 in response to NGF promotes microtubule assembly and that these factors are therefore key regulators of neurite outgrowth.

scholarly journals Regulation of nerve growth factor synthesis and release in organ cultures of rat iris.

1984 ◽  
Vol 99 (3) ◽  
pp. 839-843 ◽  
Author(s):  
E M Barth ◽  
S Korsching ◽  
H Thoenen
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Golgi Apparatus ◽  
Messenger Rna ◽  
Time Course ◽  
Maximal Level ◽  
Nerve Growth ◽  
Rat Iris

We studied the synthesis and release of nerve growth factor (NGF) in cultured rat iris with a two-site enzyme immunoassay by measuring the time course of NGF levels remaining in the iris and relased into the medium up to 72 h. For up to 3 h, the NGF levels in the iris did not change significantly. After that, they increased to a maximal level of 350 +/- 30 pg NGF/iris at 19 h, which is 200 times higher than the in vivo content. Between 20 and 72 h in culture, the NGF level decreased to 130 +/- 10 pg NGF/iris, whereas general protein synthesis did not change during that time period. Maximal rate of NGF production (203 pg NGF/h/iris) was seen between 9 and 12 h in culture. In the medium, NGF levels were first detectable after 6 h. Levels then increased with a time course similar to that seen within the iris, reaching a maximal level of 1,180 +/- 180 pg after 19 h in vitro, and then did not significantly change for up to 48 h. The NGF production of the densely sympathetically innervated dilator was three times higher than that of the predominantly cholinergically innervated sphincter. The NGF production was blocked by inhibitors of messenger RNA synthesis (actinomycin D) and of polyadenylation (9-beta-D-arabinofuranosyladenine) as well as by inhibitors of translation (cycloheximide). Monensin, which interferes with the transport of proteins through the Golgi apparatus, decreased NGF levels to 8-12% of controls in the medium, suggesting that the Golgi apparatus is involved in the intracellular processing of NGF.

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.

scholarly journals Inhibitors of protein synthesis and RNA synthesis prevent neuronal death caused by nerve growth factor deprivation.

1988 ◽  
Vol 106 (3) ◽  
pp. 829-844 ◽  
Author(s):  
D P Martin ◽  
R E Schmidt ◽  
P S DiStefano ◽  
O H Lowry ◽  
J G Carter ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neuronal Death ◽  
Adenylate Kinase ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Sympathetic Neurons ◽  
Ultrastructural Changes ◽  
Nerve Growth

We have developed an experimental paradigm to study the mechanism by which nerve growth factor (NGF) allows the survival of sympathetic neurons. Dissociated sympathetic neurons from embryonic day-21 rats were grown in vitro for 7 d in the presence of NGF. Neurons were then deprived of trophic support by adding anti-NGF antiserum, causing them to die between 24 and 48 h later. Ultrastructural changes included disruption of neurites, followed by cell body changes characterized by an accumulation of lipid droplets, changes in the nuclear membrane, and dilation of the rough endoplasmic reticulum. No primary alterations of mitochondria or lysosomes were observed. The death of NGF-deprived neurons was characterized biochemically by assessing [35S]methionine incorporation into TCA precipitable protein and by measuring the release of the cytosolic enzyme adenylate kinase into the culture medium. Methionine incorporation began to decrease approximately 18 h post-deprivation and was maximally depressed by 36 h. Adenylate kinase began to appear in the culture medium approximately 30 h after deprivation, reaching a maximum by 54 h. The death of NGF-deprived neurons was entirely prevented by inhibiting protein or RNA synthesis. Cycloheximide, puromycin, anisomycin, actinomycin-D, and dichlorobenzimidazole riboside all prevented neuronal death subsequent to NGF deprivation as assessed by the above morphologic and biochemical criteria. The fact that sympathetic neurons must synthesize protein and RNA to die when deprived of NGF indicates that NGF, and presumably other neurotrophic factors, maintains neuronal survival by suppressing an endogenous, active death program.

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.

Sign in / Sign up

Export Citation Format

Share Document