Dynamics of behaviour during neuronal morphogenesis in culture

Jacobs, J. Roger; Stevens, J. R.

doi:10.1002/cm.970080306

Cited by 12 publications

(5 citation statements)

References 46 publications

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“…PC-12 cells are highly labile in morphology shortly after NGF activation (29). More dynamic microtubules may permit rapid and reversible changes which contribute to the neurons's ability to reorganize during active growth.…”

Section: Progressive Stabilization Of Microtubules During Differentiamentioning

confidence: 99%

“…In vivo, neuritic elongation diminishes as relatively permanent connectivity between nerve cells is required for maintenance of neuronal function. Similarly, in PC-12 cells the gross morphology becomes more stable after 1 wk in NGF (29). Stable microtubules may be an important intracellular factor that constrain the capacity of the neuron to change its morphology, facilitating the maintenance of a preferred organization.…”

Section: Progressive Stabilization Of Microtubules During Differentiamentioning

confidence: 99%

“…These results are consistent with the behavior of neurons as seen in culture. Activity at the growth cones is often apparent while the neurite shaft remains relatively quiescent (1, 15,16,29). Repeated movement is observed as growth cones advance, divide, retract, and start again until a particular contact becomes stable.…”

Section: Microtubule Dynamics and Intraceuular Locationmentioning

confidence: 99%

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Progressive and spatially differentiated stability of microtubules in developing neuronal cells.

Lim

Sammak

Borisy

1989

The Journal of Cell Biology

165

104

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Abstract. The establishment of neural circuits requires both stable and plastic properties in the neuronal cytoskeleton. In this study we show that properties of stability and lability reside in microtubules and these are governed by cellular differentiation and intracellular location. After culture for 3, 7, and 14 d in nerve growth factor-containing medium, PC-12 cells were microinjected with X-rhodamine-labeled tubulin. 8-24 h later, cells were photobleached with a laser microbe.am at the cell body, neurite shaft, and growth cone. Replacement of fluorescence in bleached zones was monitored by digital video microscopy. In 3-d cultures, fluorescence recovery in all regions occurred by 26 + 17 min. Similarly, in older cultures, complete fluorescence recovery at the cell body and growth cone occurred by 10-30 min. However, in neurite shafts, fluorescence recovery was markedly slower (71 + 48 min for 7-d and 201 + 94 min for 14-d cultures). This progressive increase in the stability of microtubules in the neurite shafts correlated with an increase of acetylated microtubules. Acetylated microtubules were present specifically in the neurite shaft and not in the regions of fast microtubule turnover, the cell body and growth cone. During the recovery of fluorescence, bleached zones did not move with respect to the cell body. We conclude that the microtubule component of the neuronal cytoskeleton is differentially dynamic but stationary. DVELOPMENT of the nervous system requires that the neuronal cytoskeleton be endowed with the seemingly incompatible properties, morphological stability and plasticity. Morphological stability is critical for maintenance and proper function of neuronal circuits, while plasticity is required for cell growth and remodeling of cell processes during response to environmental inputs or to injury. How does the cytoskeleton adapt to these different requirements? Are there changes in the cytoskeleton that reflect the state of cellular differentiation?Microtubule formation is required for the growth and structural integrity of neuronal processes (17,18,62,63) and numerous studies have demonstrated that neuronal microtubules exhibit unusual stability properties. For example, a significant fraction of tubulin in brain is not solubilized by conventional methods of temperature-reversible, assembly-dissembly procedures (10,13,27,50,59). Further, the susceptibility of microtubules to depolymerization by colchicine diminishes with increased culture age (6, 17).Several factors have been postulated to enhance microtubule stability. By binding microtubule-associated proteins microtubule stability increases (4, 40), and neurons contain a variety of microtubule-associated proteins of which MAP-1, MAP-2, tau, and chartins are quite prominent (5,8,9,42,44,45). Neuronal tubulin also undergoes posttranslational modifications including detyrosination/tyrosination (2, 49), acetylation (7), and phosphorylation (21,22). Although the physiological relevance of the differential binding reactions with microtubule-asso...

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Section: Progressive Stabilization Of Microtubules During Differentiamentioning

confidence: 99%

Section: Progressive Stabilization Of Microtubules During Differentiamentioning

confidence: 99%

Section: Microtubule Dynamics and Intraceuular Locationmentioning

confidence: 99%

See 1 more Smart Citation

Progressive and spatially differentiated stability of microtubules in developing neuronal cells.

Lim

Sammak

Borisy

1989

The Journal of Cell Biology

165

104

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“…The most detailed information stems from studies with the neuronlike PC12 model system, which can be induced to extend neurites by treatment with NGF (Greene and Tischler, 1976). In cultures of PC12 cells, as well as of neurons, early neurite outgrowth is characterized by the initiation, elongation, and then retraction of neurites (Jacobs and Stevens, 1987). Eventually, the behavior of the cell stabilizes so that specific neurites persist and continue to elongate.…”

Section: Microtubule Dynamics In Neuronal Versus Non-neuronal Cellsmentioning

confidence: 99%

Dynamics of alpha-tubulin deacetylation in intact neurons

1989

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The majority of the alpha-tubulin in cultured neurons is acetylated (Black and Keyser, 1987). The present studies examine the relationships of the acetylation and deacetylation reactions to tubulin assembly and disassembly in intact neurons. Extraction assays which separate assembled and unassembled tubulin pools reveal that greater than or equal to 99% of the total acetylated, as well as newly acetylated, tubulin is cytoskeletal associated. Treatment of neurons with depolymerizing drugs results in a progressive decrease in the levels of total tubulin in polymer and a corresponding increase in the levels of soluble tubulin. These drugs also cause a progressive decrease in the levels of acetylated alpha-tubulin in polymer that closely parallels in rate and extent that of total alpha-tubulin. However, there is no corresponding increase in soluble acetylated tubulin. Because the total levels of alpha-tubulin remain unchanged during drug treatment, the decrease in levels of acetylated alpha-tubulin during depolymerization must reflect its rapid conversion to nonacetylated alpha-tubulin. These findings suggest alpha-tubulin is acetylated in the polymeric form and that deacetylation is closely coupled to depolymerization. The close coupling between alpha-tubulin deacetylation and depolymerization provided a means of estimating the rate at which subunits cycle off microtubules in intact neurons. Acetate turnover on tubulin in intact neurons was determined both by pulse-chase protocols with 3H-acetate and by measuring the loss of acetylated subunits (using quantitative immunoblotting) under conditions of net microtubule depolymerization induced by colchicine. Both methods yielded similar results. Acetate turnover occurred biphasically; 30-50% of the acetate on tubulin turns over with a t1/2 of 1.5-2 hr, and the remaining half or more turns over with a t1/2 of 5-10 hr. We suggest that these kinetically distinguishable pools of acetylated alpha-tubulin reflect distinct pools of acetylated microtubules that differ in their average rates of subunit turnover.

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“…Neuronal differentiation refers to the physiological, biochemical and morphological changes that cells undergo in order to become mature neurons [24] . Neurotrophins such as nerve growth factor (NGF) have been shown to induce PC12 cell differentiation characterized by cell cycle arrest, neurite outgrowth and the emergence of differentiation markers [25,26] .…”

Section: Introductionmentioning

confidence: 99%

Heparin-Binding Epidermal Growth Factor-Like Growth Factor Is a Potent Neurotrophic Factor for PC12 Cells

Zhou

Besner

2010

Neurosignals

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Heparin-binding epidermal growth factor EGF-like growth factor (HB-EGF) is a member of the epidermal growth factor family that is expressed in many cell types. We have previously reported the effects of HB-EGF on intestinal epithelial cells and endothelial cells after exposure to ischemia/reperfusion in vivo or anoxia/reoxygenation injury in vitro. However, the effect of HB-EGF on neuronal cells is largely unexplored. In this study, we examined the effect of HB-EGF on neurite outgrowth in pheochromocytoma (PC12) cells as well as the neuroprotective effect of HB-EGF on injured PC12 cells exposed to oxygen and glucose deprivation (OGD), which mimics ischemic conditions. We found that HB-EGF significantly promotes PC12 cell neurite outgrowth and that this effect was blocked by EGF receptor (EGFR) inhibition or mitogen-activated protein kinase (MAPK) inhibition, but not by tyrosine kinase inhibition. In the face of OGD injury, HB-EGF preserves cell viability and decreases apoptosis and LDH release in PC12 cells. HB-EGF-mediated cytoprotection was abolished by EGFR inhibition and MAPK inhibition. We conclude that HB-EGF, through its interaction with the EGF receptor, activates the MAPK signaling pathway in PC12 cells under basal or injury conditions, leading to enhanced neurite outgrowth and neuroprotection against ischemic injury.

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Dynamics of behaviour during neuronal morphogenesis in culture

Cited by 12 publications

References 46 publications

Progressive and spatially differentiated stability of microtubules in developing neuronal cells.

Progressive and spatially differentiated stability of microtubules in developing neuronal cells.

Dynamics of alpha-tubulin deacetylation in intact neurons

Heparin-Binding Epidermal Growth Factor-Like Growth Factor Is a Potent Neurotrophic Factor for PC12 Cells

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