Density-dependent nerve growth factor regulation of Gs-alpha RNA in pheochromocytoma 12 cells.

Nerve growth factor (NGF) affects levels of the alpha subunit of the stimulatory G protein (Gs-alpha) in pheochromocytoma 12 cells in a bidirectional, density-dependent manner. Cells grown at high density responded to NGF treatment with increased levels of Gs-alpha mRNA and protein. Conversely, in cells grown in low-density cultures, levels of this mRNA were lowered by NGF treatment.

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Mouse 7S nerve growth factor: complete sequence of a cDNA coding for the .alpha.-subunit precursor and its relationship to serine proteases

Biochemistry ◽

10.1021/bi00320a015 ◽

1984 ◽

Vol 23 (25) ◽

pp. 5997-6002 ◽

Cited By ~ 39

Author(s):

Paul J. Isackson ◽

Axel Ullrich ◽

Ralph A. Bradshaw

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Serine Proteases ◽

Complete Sequence ◽

Alpha Subunit ◽

Nerve Growth

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Nerve Growth Factor Stimulates Multisite Tyrosine Phosphorylation and Activation of the Atypical Protein Kinase C's via a src Kinase Pathway

Molecular and Cellular Biology ◽

10.1128/mcb.21.24.8414-8427.2001 ◽

2001 ◽

Vol 21 (24) ◽

pp. 8414-8427 ◽

Cited By ~ 66

Author(s):

Marie W. Wooten ◽

Michel L. Vandenplas ◽

M. Lamar Seibenhener ◽

Thangiah Geetha ◽

Maria T. Diaz-Meco

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Protein Kinase ◽

Tyrosine Phosphorylation ◽

Pc12 Cells ◽

Catalytic Domain ◽

Src Kinase ◽

Dependent Manner ◽

Nerve Growth ◽

Atypical Protein Kinase

ABSTRACT Atypical protein kinase C (PKC) isoforms are required for nerve growth factor (NGF)-initiated differentiation of PC12 cells. In the present study, we report that PKC-ι becomes tyrosine phosphorylated in the membrane coincident with activation posttreatment with nerve growth factor. Tyrosine phosphorylation and activation of PKC-ι were inhibited in a dose-dependent manner by both PP2 and K252a, src and TrkA kinase inhibitors. Purified src was observed to phosphorylate and activate PKC-ι in vitro. In PC12 cells deficient in src kinase activity, both NGF-induced tyrosine phosphorylation and activation of PKC-ι were also diminished. Furthermore, we demonstrate activation of src by NGF along with formation of a signal complex including the TrkA receptor, src, and PKC-ι. Recruitment of PKC-ι into the complex was dependent on the tyrosine phosphorylation state of PKC-ι. The association of src and PKC-ι was constitutive but was enhanced by NGF treatment, with the src homology 3 domain interacting with a PXXP sequence within the regulatory domain of PKC-ι (amino acids 98 to 114). Altogether, these findings support a role for src in regulation of PKC-ι. Tyrosine 256, 271, and 325 were identified as major sites phosphorylated by src in the catalytic domain. Y256F and Y271F mutations did not alter src-induced activation of PKC-ι, whereas the Y325F mutation significantly reduced src-induced activation of PKC-ι. The functional relevance of these mutations was tested by determining the ability of each mutant to support TRAF6 activation of NF-κB, with significant impairment by the Y325F PKC-ι mutant. Moreover, when the Y352F mutant was expressed in PC12 cells, NGF's ability to promote survival in serum-free media was reduced. In summary, we have identified a novel mechanism for NGF-induced activation of atypical PKC involving tyrosine phosphorylation by c-Src.

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Spatial Distribution of Nociceptive Neuropeptide and Nerve Growth Factor Depletion in Experimental Diabetic Peripheral Nervous System

Journal of International Medical Research ◽

10.1177/147323000203000507 ◽

2002 ◽

Vol 30 (5) ◽

pp. 512-519 ◽

Cited By ~ 6

Author(s):

H Kanbayashi ◽

H Itoh ◽

T Kashiwaya ◽

K Atoh ◽

I Makino

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Substance P ◽

Dorsal Root ◽

Time Course ◽

Diabetic Control ◽

Diabetic Rats ◽

Enzyme Linked Immunosorbent Assay ◽

Dependent Manner ◽

Nerve Growth

This study investigated the time-course of the nociceptive neuropeptide substance P and nerve growth factor (NGF), which facilitates substance P production, in lumbar and cervical dorsal root ganglia (DRG) of streptozotocin-induced diabetic rats. Levels of substance P and NGF were measured by radioimmunoassay and sandwich enzyme-linked immunosorbent assay, respectively, 2 months, 4 months and 8 months after induction of diabetes, and compared with age-matched non-diabetic control rats. At 2 months and 4 months, substance P and NGF levels were lower in the lumbar DRG of the diabetic rats than in controls. At 8 months, substance P and NGF were lower in both the lumbar and cervical DRG of the diabetic rats than in controls. These data demonstrate that a decrease in substance P levels in primary sensory neurons with NGF depletion occurs in an axonal length-dependent manner in diabetic rats, and that this decrease may be correlated with the duration of diabetes.

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Reduced Number, G Protein Coupling, and Antinociceptive Efficacy of Spinal Mu-Opioid Receptors in Diabetic Rats Are Reversed by Nerve Growth Factor

Journal of Pain ◽

10.1016/j.jpain.2013.01.776 ◽

2013 ◽

Vol 14 (7) ◽

pp. 720-730 ◽

Cited By ~ 21

Author(s):

Mohammed Shaqura ◽

Baled I. Khalefa ◽

Mehdi Shakibaei ◽

Jens Winkler ◽

Mahmoud Al-Khrasani ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

G Protein ◽

Opioid Receptors ◽

Diabetic Rats ◽

Mu Opioid Receptors ◽

G Protein Coupling ◽

Nerve Growth ◽

Mu Opioid ◽

Protein Coupling

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Phosphorylation of translation initiation factor eIF-4E is induced in a ras-dependent manner during nerve growth factor-mediated PC12 cell differentiation.

Molecular and Cellular Biology ◽

10.1128/mcb.12.3.1239 ◽

1992 ◽

Vol 12 (3) ◽

pp. 1239-1247 ◽

Cited By ~ 63

Author(s):

R M Frederickson ◽

W E Mushynski ◽

N Sonenberg

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Pc12 Cells ◽

Translation Initiation ◽

Sympathetic Neurons ◽

Translation Initiation Factor ◽

Tumor Promoter ◽

Initiation Factor ◽

Dependent Manner ◽

Nerve Growth

Translation initiation factor eIF-4E, which binds to the 5' cap structure of eukaryotic mRNAs, is believed to play an important role in the control of cell growth. Consistent with this, overexpression of eIF-4E in fibroblasts results in their malignant transformation. The activity of eIF-4E is thought to be regulated by phosphorylation on a single serine residue (Ser-53). Treatment of rat pheochromocytoma (PC12) cells with nerve growth factor (NGF) strongly curtails their growth and causes their differentiation into cells that resemble sympathetic neurons. The present study shows that eIF-4E is rapidly phosphorylated in PC12 cells upon NGF treatment, resulting in a significant increase in the steady-state levels of the phosphorylated protein. In contrast, epidermal growth factor, a factor which elicits a weak mitogenic response in PC12 cells, did not significantly enhance eIF-4E phosphorylation. We also show that although the mitogen and tumor promoter, phorbol 12-myristate-13-acetate, is able to induce phosphorylation of eIF-4E in PC12 cells, the NGF-mediated increase is primarily a protein kinase C-independent response. The NGF-induced enhancement of eIF-4E phosphorylation is abrogated in PC12 cells expressing a dominant inhibitory ras mutant (Ser-17 replaced by Asn), indicating that eIF-4E phosphorylation is dependent on a ras signalling pathway. As phosphorylation of eIF-4E effects translation initiation, these results suggest that NGF-mediated and ras-dependent eIF-4E phosphorylation may play a role in switching the pattern of gene expression during the differentiation of PC12 cells.

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Impaired production of nerve growth factor in the submandibular gland of diabetic mice

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1989.257.3.e400 ◽

1989 ◽

Vol 257 (3) ◽

pp. E400-E404 ◽

Cited By ~ 4

Author(s):

S. Kasayama ◽

T. Oka

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Submandibular Gland ◽

Control Level ◽

Molecular Size ◽

Dependent Manner ◽

Diabetic Mice ◽

Western Blots ◽

Nerve Growth ◽

Do So

The production of nerve growth factor (NGF) in submandibular glands was examined in two kinds of diabetic mice. In genetically diabetic (C57BL/KsJ db/db) mice, which manifest marked insulin resistance and hyperglycemia, the concentration of NGF in the submandibular gland was less than one-tenth that of the nondiabetic controls. In streptozotocin-induced diabetic C57BL/KsJ mice, which show pancreatic insulitis leading to insulin deficiency and hyperglycemia, the glandular NGF concentration fell in a time-dependent manner to 26% of control level at 5 wk after the streptozotocin injection. A daily administration of insulin to the streptozotocin-induced diabetic mice restored the NGF concentration to almost the control level. The molecular size of NGF (13 kDa) in the glandular extracts of the genetically diabetic (db/db) mice in Western blots was indistinguishable from that of the control mice, but its level was reduced in the glands of the diabetic (db/db) animals. Although plasma NGF concentrations were normally below the sensitivity of the assay (less than 0.80 ng/ml) in both the control and the diabetic (db/db) mice, administration of cyclocytidine, which stimulates NGF release from the submandibular gland into the blood circulation, increased the plasma NGF level to 5.95 ng/ml in the control mice, but it failed to do so in the diabetic (db/db) mice. These findings suggest that, in diabetic mice, NGF production in the submandibular gland and its capacity to release NGF into the circulation are decreased.

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Oncogene N-ras mediates selective inhibition of c-fos induction by nerve growth factor and basic fibroblast growth factor in a PC12 cell line.

Molecular and Cellular Biology ◽

10.1128/mcb.10.4.1556 ◽

1990 ◽

Vol 10 (4) ◽

pp. 1556-1563 ◽

Cited By ~ 16

Author(s):

T M Thomson ◽

S H Green ◽

R J Trotta ◽

D E Burstein ◽

A Pellicer

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Cell Line ◽

Basic Fibroblast Growth Factor ◽

Dependent Manner ◽

Fibroblast Growth ◽

Induced Expression ◽

Nerve Growth ◽

Ras P21

A cell line was generated from U7 cells (a subline of PC12 rat pheochromocytoma cells) that contains a stably integrated transforming mouse N-ras (Lys-61) gene under the control of the long terminal repeat from mouse mammary tumor virus. Such cells, designated UR61, undergo neuronal differentiation upon exposure to nanomolar concentrations of dexamethasone, as a consequence of expression of the activated N-ras gene (I. Guerrero, A. Pellicer, and D.E. Burstein, Biochem, Biophys. Res. Commun. 150:1185-1192, 1988). Exposure of UR61 cells to either nerve growth factor (NGF) or basic fibroblast growth factor (bFGF) results in a marked induction of c-fos RNA, with kinetics paralleling those of NGF- or bFGF-induced expression of c-fos RNA in PC12 cells. Dexamethasone-induced expression of activated N-ras p21 results in blocking of c-fos RNA induction by NGF or bFGF in a time-dependent manner. Activated N-ras p21-mediated inhibition of c-fos RNA induction in UR61 cells is selective for NGF and bFGF and is not due to selective degradation of c-fos RNA. Normal and transforming N-ras can trans activate the chloramphenicol acetyltransferase gene linked to mouse c-fos regulatory sequences when transient expression assays are performed. Our observations suggest that N-ras p21 selectively interacts with pathways involved in induction of c-fos expression which initiate at the receptors for NGF and bFGF.

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Exploring Human Testicular Peritubular Cells: Identification of Secretory Products and Regulation by Tumor Necrosis Factor-α

Endocrinology ◽

10.1210/en.2007-1064 ◽

2008 ◽

Vol 149 (4) ◽

pp. 1678-1686 ◽

Cited By ~ 68

Author(s):

Christoph Schell ◽

Martin Albrecht ◽

Christine Mayer ◽

J. Ullrich Schwarzer ◽

Monica B. Frungieri ◽

...

Keyword(s):

Nerve Growth Factor ◽

Growth Factor ◽

Cyclooxygenase 2 ◽

Time Dependent ◽

Human Testis ◽

Dependent Manner ◽

Phenotypic Characteristics ◽

Nerve Growth ◽

Tnf Α ◽

Peritubular Cells

Testicular peritubular cells are myofibroblastic cells, which represent the major cellular components of the wall of the seminiferous tubules. In men their phenotypic characteristics, including possible secretory activity and regulation, are not well known, in neither normal nor pathologically altered testes. Especially in testes of men with impaired spermatogenesis, the cytoarchitecture of the tubular wall is frequently remodeled and presents fibrotic thickening, increased innervation, and infiltration by macrophages and mast cells. The latter are two sources of TNF-α. The purpose of our study was to explore human testicular peritubular cells and mechanisms of their regulation. To this end we primarily studied cultured human testicular peritubular cells (HTPCs), isolated from adult human testes. Having established that HTPCs express TNF-α receptors 1 and 2 and respond to recombinant human TNF-α by a rapid phosphorylation of ERK1/2, we used complementary approaches, including gene array/RT-PCR studies, Western blotting/immunocytochemistry, and ELISA techniques to study phenotypic characteristics of HTPCs and actions of TNFα. We found that HTPCs express the nerve growth factor gene and TNF-α-stimulated mRNA levels and secretion of nerve growth factor in a dose- and time-dependent manner. Similarly, monocyte chemoattractant protein-1 was identified as a product of HTPCs, which was regulated by TNF-α in a concentration- and time-dependent manner. TNF-α furthermore strongly enhanced expression and/or synthesis of other inflammatory molecules, namely IL-6 and cyclooxygenase-2. Active cyclooxygenase-2 is indicated by increased prostaglandin D2 levels. In addition, intercellular adhesion molecule-1, which was not detected at protein level in the absence of TNF-α, was induced upon TNF-α stimulation. In conclusion, these results provide novel insights into the nature of human peritubular cells, which are able to secrete potent signaling molecules and are regulated by TNF-α. These results also hint to an as-yet-unknown role of peritubular cells in normal human testis and involvement in the pathomechanisms associated with impaired spermatogenesis in men.

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