Nerve Growth Factor Uses Ras/ERK and Phosphatidylinositol 3-Kinase Cascades to Up-regulate theN-Methyl-d-aspartate Receptor 1 Promoter

Liu, Anguo; Prenger, Michael S.; Norton, Darrell D.; Mei, Lin; Kusiak, John W.; Bai, Guang

doi:10.1074/jbc.m105399200

Cited by 48 publications

(52 citation statements)

References 65 publications

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“…Luciferase activity was normalized to ␤-galactosidase activity. Sp factor constructs were described in previous studies (39).…”

Section: Methodsmentioning

confidence: 99%

“…DNA constructs of the NR2A promoter luciferase gene and pCMV␤gal (9:1) were transfected to cells. Two days after transfection, cells were lysed for reporter gene assays as described previously (39). Transfection efficiency was about 5% determined by green fluorescent cells after transfection of pEGFP (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

“…DNA products were visualized with UV illumination on agarose gel containing 10 g/ml ethidium bromide. Images were taken on a Kodak Image Station 440CF and analyzed with 1D Image Analysis software as stated previously (39).…”

Section: Methodsmentioning

confidence: 99%

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Functional Analysis of the Rat N-Methyl-D-aspartate Receptor 2A Promoter

Liu

Zhuang

Hoffman

et al. 2003

Journal of Biological Chemistry

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N-Methyl-D-aspartate (NMDA) receptor subunit 2A (NR2A) is an important modulatory component of the NMDA subtype of glutamate receptors. To investigate the transcription mechanism of the NR2A gene, we cloned the 5 -flanking sequence from a rat genomic library. RNA mapping with rat brain RNA revealed two sets of major and several minor transcription start points in a single exon of 1140 bp. Reporter gene and mutation studies indicated that core promoter activity resided in exon 1, whereas the 5 -flanking sequence up to 1.5 kb showed no significant impact on promoter activity. Fragments containing minor transcription start points were able to drive a reporter gene in transfected cells and produce nascent RNAs in an in vitro transcription system. All fragments tested showed more promoter activity in dissociated neurons of the rat embryonic cerebrocortex and cell lines expressing NR2A mRNA than that in glial cultures and non-neuronal cells. Within exon 1 there are three GC-box elements that displayed distinct binding affinity to both Sp1-and Sp4-like factors. Overexpression of Sp1 or Sp4, but not Sp3, significantly increased the activity of the promoter containing these elements. Inclusion of exon 2 and 3 sequences, which contain five short open-reading frames, attenuated promoter-driven reporter activity more than 3-fold but attenuated the level of reporter mRNA less than 1.4-fold. Our results suggest that the core promoter of the rat NR2A gene requires exon 1, that Sp factors positively regulate this core promoter, and that a post-transcriptional mechanism may negatively regulate expression of the gene.

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“…Luciferase activity was normalized to ␤-galactosidase activity. Sp factor constructs were described in previous studies (39).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Functional Analysis of the Rat N-Methyl-D-aspartate Receptor 2A Promoter

Liu

Zhuang

Hoffman

et al. 2003

Journal of Biological Chemistry

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“…Also, it is reported that NGF is mediated by PI3K/Akt/mTOR pathway [281][282][283]. Ras-ERK pathway is a key promoter of neurite outgrowth and also has been found to enhance axonal survival [284]. Rho-ROK pathway participates in neural growth and modulates neurite outgrowth [285].…”

Section: Pharmacological Agentsmentioning

confidence: 99%

Scaffolds for Peripheral Nerve Regeneration, the Importance of In Vitro and In Vivo Studies for the Development of Cell-Based Therapies and Biomaterials: State of the Art

Pedrosa¹,

Caseiro²,

Santos³

et al. 2017

Scaffolds in Tissue Engineering - Materials, Technologies and Clinical Applications

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Human adult peripheral nerve injuries are a high incidence clinical problem that greatly affects patients' quality of life. Although peripheral nervous system has intrinsic regenerative capacity, this occurs in an incomplete or poorly functional manner. When a nerve fiber loses its continuity with consequent damage of the basal lamina tubes, axon spontaneous regeneration is disorganized and mismatched. These phenomena translate in an inadequate nerve functional recovery and consequent musculoskeletal incapacity. Nerve grafts still remain the gold standard in peripheral injuries treatment. However, this approach contains its disadvantages such as the necessity of primary surgery to harvest the autografts, loss of a functional nerve, donor site morbidity and longer surgery procedures. Therefore, biomaterials and tissue engineering can provide efficient resources and alternatives to nerve injury repair not only by the development of biocompatible structures but also, introducing neurotrophic factors and cellular systems to stimulate optimum clinical outcome. In this chapter, a comprehensive state-of-the art picture of tissue-engineered nerve grafts scaffolds, their application in nerve regeneration along with latest advances in peripheral nerve repair and future perspectives will be discussed, including our own large experience in this field of knowledge.

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“…The zinc finger protein Sp1 participates in the transcriptional regulation of several other NGF-responsive genes (Liu et al, 2001;Melnikova and Gardner, 2001). Functional Sp1 binding elements are located at Ϫ48 and Ϫ168 bp in the rat PNMT promoter , and we have shown that the Ϫ48 bp Sp1 site contributes to the cooperative activation of the PNMT promoter by PKA and PKC .…”

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confidence: 99%

Nerve Growth Factor Regulates Adrenergic Expression

Tai

Wong-Faull²,

Claycomb³

et al. 2006

Mol Pharmacol

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The mechanism by which nerve growth factor (NGF) regulates adrenergic expression was examined in PC-12 cells transfected with a rat phenylethanolamine N-methyl-transferase (PNMT) promoter-luciferase reporter gene construct pGL3RP893. NGF treatment increased PNMT promoter-driven luciferase activity in a dose-and time-dependent manner. Induction was attenuated by inhibition of the extracellular signalregulated kinase mitogen-activated protein kinase (MAPK) pathway (ϳ60%) but not by inhibition of the protein kinase A (PKA), protein kinase C, phosphoinositol kinase, or p38 MAPK pathways. Deletion PNMT promoter-luciferase reporter gene constructs showed that the NGF-responsive sequences lay within the proximal Ϫ392 base pairs (bp) of PNMT promoter, wherein binding elements for Egr-1 (Ϫ165 bp) and Sp1 (Ϫ48 bp) reside. Western analysis further showed that NGF increased nuclear levels of Egr-1, but not Sp1 or the catalytic subunit of PKA. Gel mobility shift assays showed increased potential for Egr-1, but not Sp1, protein-DNA binding complex formation. Mutation of either the Egr-1 or Sp1 binding sites in the PNMT promoter attenuated NGF activation. NGF, combined with pituitary adenylyl cyclase-activating protein (PACAP), another PNMT transcriptional activator, cooperatively stimulated PNMT promoter driven-luciferase activity beyond levels observed with either neurotrophin alone. Finally, posttranscriptional control seems to be another important mechanism by which neurotrophins regulate the adrenergic phenotype. NGF, PACAP, and a combination of the two stimulated both intron-retaining and intronless PNMT mRNA and PNMT protein, but to different extents.

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Nerve Growth Factor Uses Ras/ERK and Phosphatidylinositol 3-Kinase Cascades to Up-regulate theN-Methyl-d-aspartate Receptor 1 Promoter

Cited by 48 publications

References 65 publications

Functional Analysis of the Rat N-Methyl-D-aspartate Receptor 2A Promoter

Functional Analysis of the Rat N-Methyl-D-aspartate Receptor 2A Promoter

Scaffolds for Peripheral Nerve Regeneration, the Importance of In Vitro and In Vivo Studies for the Development of Cell-Based Therapies and Biomaterials: State of the Art

Nerve Growth Factor Regulates Adrenergic Expression

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