Nerve growth factor prevents the death and stimulates the neuronal differentiation of clonal PC12 pheochromocytoma cells in serum-free medium

Greene, LA

doi:10.1083/jcb.78.3.747

Cited by 432 publications

(373 citation statements)

References 30 publications

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“…The amount of protein and the amount of total RNA per dish was equivalent for comparisons of NGFtreated and untreated cells cultured for 5 days. However, the cell number of the NGF-treated cultures was significantly lower (2.6 ϫ 10 7 cells in untreated 100-mm-diameter dishes compared to 0.83 ϫ 10 7 cells in those exposed to NGF), consistent with the known functions of NGF to halt proliferation, trigger the differentiation, and support survival of PC12 cells (22,23,50). Whereas the relative amount of CCT␤2 transcript per cell increased by a factor of 3.8, from 0.268 to 1.02 in our measurements, in agreement with the previous report (12), the amount of CCT␣ transcript per cell also increased, from 1.0 to 2.48 (Fig.…”

Section: Fig 3 Expression Of Cct␣ and Cct␤ Mrnas In Tissues Frommentioning

confidence: 53%

Disruption of CCTβ2 Expression Leads to Gonadal Dysfunction

Jackowski

Rehg

Zhang³

et al. 2004

Molecular and Cellular Biology

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There are two mammalian genes that encode isoforms of CTP:phosphocholine cytidylyltransferase (CCT), a key rate-controlling step in membrane phospholipid biogenesis. Quantitative determination of the CCT transcripts reveals that CCT␣ is ubiquitously expressed and is found at the highest levels in the testis and lung, with lower levels in the liver and ovary. CCT␤2 is a very minor isoform in most tissues but is significantly expressed in the brain, lung, and gonads. CCT␤3 is the third isoform recently discovered in mice and is expressed in the same tissues as CCT␤2, with its highest level in testes. We investigated the role(s) of CCT␤2 by generating knockout mice. The brains and lungs of mice lacking CCT␤2 expression did not exhibit any overt defects. On the other hand, a large percentage of the CCT␤2 ؊/؊ females were sterile and their ovaries exhibited defective ovarian follicle development. The proportion of female CCT␤2 ؊/؊ mice with defective ovaries increased as the animals aged. The rare litters born from CCT␤2 ؊/؊ ؋ CCT␤2 ؊/0 matings had the normal number of pups. The abnormal ovarian histopathology was characterized by disorganization of the tissue in young adult mice and absence of follicles and ova in older mice, along with interstitial stromal cell hyperplasia which culminated in the emergence of tubulostromal ovarian tumors by 16 months of age. Grossly defective CCT␤2 ؊/؊ ovaries were associated with high follicle-stimulating (FSH) and luteinizing (LH) hormone levels. Male CCT␤2 ؊/0 mice exhibited progressive multifocal testicular degeneration and reduced fertility but had normal FSH and LH levels. Thus, the most notable phenotype of CCT␤2 knockout mice was gonad degeneration and reproductive deficiency. The results indicate that although CCT␤2 is expressed at very low levels compared to the ␣-isoform, loss of CCT␤2 expression causes a breakdown in the gonadal response to hormonal stimulation.Phosphatidylcholine (PtdCho) is a major component of biological membranes in higher eukaryotes and is also secreted by specialized tissues for important extracellular tasks. CTP: phosphocholine cytidylyltransferase (CCT) is a key rate-controlling step in the major biosynthetic pathway leading to PtdCho in most tissues (for reviews, see references 15, 17, and 36). In mammals there are two genes, Pcyt1a (formerly Cptct), located on murine chromosome 16, and Pcyt1b, located on the X chromosome, that encode proteins termed CCT␣ and CCT␤, respectively (35, 38). The two genes exhibit tissuespecific expression, with CCT␣ predominating in most tissues and CCT␤ being most abundant in brain tissue (32). Two transcripts arise from the Pcyt1a gene that encode the identical CCT␣ protein. Alternate splicing of the X-linked Pcyt1b gene directs the synthesis of two mRNAs that encode the CCT␤2 and CCT␤3 isoforms in mice (32,35,38). CCT␤3 is 28 residues shorter at the amino terminus than CCT␤2 due to transcript initiation at an alternate first exon (32). In humans, intron retention gives rise to a CCT␤1 transcript found in the exp...

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Section: Fig 3 Expression Of Cct␣ and Cct␤ Mrnas In Tissues Frommentioning

confidence: 53%

Disruption of CCTβ2 Expression Leads to Gonadal Dysfunction

Jackowski

Rehg

Zhang³

et al. 2004

Molecular and Cellular Biology

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“…This observation is highly reminiscent of the apoptotic cell death of differentiated PC12 cells after NGF withdrawal (Greene, 1978). Similarly, the inhibition of BTG2 TIS21/PC3 expression may trigger programmed cell death as a consequence of terminally differentiated cells attempting to re-enter the cycle.…”

Section: Btg2 Tis21/pc3 Triggers Differentiation Of Pc12 Cells F El-gmentioning

confidence: 90%

BTG2TIS21/PC3 induces neuronal differentiation and prevents apoptosis of terminally differentiated PC12 cells

et al. 2002

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The p53-transcriptional target, BTG2 TIS21/PC3 , was previously identified as an antiproliferative gene. However, the precise biological functions of the protein product remain to be elucidated. BTG2 TIS21/PC3 expression is induced in vivo during neurogenesis, and the gene is transiently expressed in vitro in rat pheochromocytoma PC12 cells after induction of neuronal differentiation by addition of nerve growth factor (NGF). These observations suggest that BTG2 TIS21/PC3 is functionally significant during the neuronal differentiation process. To test this hypothesis, a vector that expressed BTG2 TIS21/PC3 under the control of an inducible promoter was introduced into PC12 cells. Growth arrest and differentiation in response to NGF were greatly enhanced by BTG2 TIS21/PC3 overexpression. Furthermore, an antisense oligonucleotide complementary to BTG2 TIS21/PC3 mRNA, which was able to inhibit endogenous BTG2 TIS21/PC3 expression, triggered programmed cell death in differentiated PC12 cells. These observations confirm that BTG2 TIS21/PC3 expression promotes neuronal differentiation and that it is required for survival of terminally differentiated cells.

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“…Removal of NGF is followed by degeneration of processes within 24 hr and by resumption of cell multiplication within 72 hr.173 NGF prevents the death and stimulates the neuronal differentiation of PC 12 cells in serum-free medium. 174 Because of their ease of handling and inducible neuron-like behavior, PC 12 cells were used to test the neuroinductive, neuroconductive, and degradative qualities of proteingraft-PEG hydrogels. In particular, the hope was that PCl2 cells in the presence of NGF would attach to, degrade, and extend neuritic processes inside protein-graff-PEG hydrogels, thereby verifying the material's intended functions in a neuronal context.…”

Section: Pc12 Cellsmentioning

confidence: 99%

Biologically Engineered Protein-graft-Poly(ethylene glycol) Hydrogels: A Cell Adhesive and Plasmin-Degradable Biosynthetic Material for Tissue Repair

et al. 2002

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To address the need for bioactive materials toward clinical applications in wound healing and tissue regeneration, an artificial protein was created by recombinant DNA methods and modified by grafting of poly(ethylene glycol) diacrylate. Subsequent photopolymerization of the acrylate-containing precursors yielded protein-graft-poly(ethylene glycol) hydrogels. The artificial protein contained repeating amino acid sequences based on fibrinogen and anti-thrombin III, comprising an RGD integrin-binding motif, two plasmin degradation sites, and a heparin-binding site. Two-dimensional adhesion studies showed that the artificial protein had specific integrin-binding capability based on the RGD motif contained in its fibrinogen-based sequence. Furthermore, heparin bound strongly to the protein's anti-thrombin III-based region. Protein-graft-poly(ethylene glycol) hydrogels were plasmin degradable, had Young's moduli up to 3.5 kPa, and supported three-dimensional outgrowth of human fibroblasts. Cell attachment in three dimensions resulted from specific cell-surface integrin binding to the material's RGD sequence. Hydrogel penetration by cells involved serine-protease mediated matrix degradation in temporal and spatial synchrony with cellular outgrowth. Protein-graft-poly(ethylene glycol) hydrogels represent a new and versatile class of biomimetic hybrid materials that hold clinical promise in serving as implants to promote wound healing and tissue regeneration.

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Nerve growth factor prevents the death and stimulates the neuronal differentiation of clonal PC12 pheochromocytoma cells in serum-free medium

Cited by 432 publications

References 30 publications

Disruption of CCTβ2 Expression Leads to Gonadal Dysfunction

Disruption of CCTβ2 Expression Leads to Gonadal Dysfunction

BTG2TIS21/PC3 induces neuronal differentiation and prevents apoptosis of terminally differentiated PC12 cells

Biologically Engineered Protein-graft-Poly(ethylene glycol) Hydrogels: A Cell Adhesive and Plasmin-Degradable Biosynthetic Material for Tissue Repair

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