Inducibility of BDNF Gene Promoter I Detected by Calcium-Phosphate-Mediated DNA Transfection Is Confined to Neuronal but Not to Glial Cells

Tabuchi, Akiko; Sano, Kuniaki; Nakaoka, Ryuki; Nakatani, Chikako; Tsuda, Masashi

doi:10.1006/bbrc.1998.9854

Cited by 29 publications

(25 citation statements)

References 10 publications

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“…Each vector was transfected with Renilla luciferase control vector driven by EF1␣ promoter (pRL-EF1␣) into rat cortical neurons at 3 DIV. We previously reported that the luciferase activity derived from the vector carrying the region from Ϫ528 to ϩ138 increased in response to the Ca 2ϩ influx through L-VDCC, which was caused by 25 mM KCl stimulation, suggesting the presence of Ca 2ϩ -responsive elements within the region up to nucleotide position Ϫ528 on BDNF-PI (18). As shown in Fig.…”

Section: Calcium-responsive Elements Located In Two Distinctmentioning

confidence: 72%

“…Cell Culture-Primary cultures of rat cortical neurons were prepared from the cerebral cortexes of 17-18-day-old rat (Sprague-Dawley) embryos as described previously (18). Briefly, small pieces of cerebral cortex were dissected by enzymatic (DNase I (Sigma) followed by trypsin (Sigma)) treatment and mechanical dissociation, and the cells were seeded at 5 ϫ 10 6 cells in a 60-mm culture dish (Iwaki).…”

Section: Methodsmentioning

confidence: 99%

“…In addition, we have also reported that BDNF-PI is activated by membrane depolarization, the basal promoter activity being ϳ8-fold less than that of BDNF-PIII (7,18). To confirm whether endogenous 5Ј-exon specific BDNF transcripts, particularly those containing exon I, are induced by membrane depolarization or not, we examined the increase in each transcript by RT-PCR.…”

Section: Production Of Bdnf Transcripts Containing Exons I Ii Iii mentioning

confidence: 99%

“…The procedure for plasmid construction was described previously (18). As an internal control vector against the luciferase experimental vector, we used Renilla luciferase vector carrying the human elongation factor 1␣ promoter (pRL-EF1␣) (7).…”

Section: Rna Isolation and Reverse Transcription (Rt)-pcr-totalmentioning

confidence: 99%

“…DNA Transfection, Luciferase, and ␤-Galactosidase Assays-DNA transfection was carried out over 3 days in culture and using calcium phosphate/DNA precipitation as already described (18). Briefly, the calcium phosphate/DNA precipitates were prepared by mixing one volume (100 l) of plasmid DNA (6 g, pBDNFpI:pRL-EF1␣ ϭ 10:1) in a 250 mM CaCl 2 solution with an equal volume of 2ϫ HEPES-buffered saline (50 mM HEPES-NaOH (pH7.05), 280 mM NaCl, 1.5 mM Na 2 HPO 4 ), and added to each 60-mm dish.…”

Section: Rna Isolation and Reverse Transcription (Rt)-pcr-totalmentioning

confidence: 99%

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Involvement of an Upstream Stimulatory Factor as Well as cAMP-responsive Element-binding Protein in the Activation of Brain-derived Neurotrophic Factor Gene Promoter I

Tabuchi¹,

Sakaya²,

Kisukeda³

et al. 2002

Journal of Biological Chemistry

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, BDNF-PI was markedly activated, as well as BDNF-PIII, by Ca 2؉ signals evoked via neuronal activity. However, little is known about the mechanisms for the transcriptional activation of BDNF-PI. Using rat cortical neurons in culture, we assigned the promoter sequences responsible for the Ca 2؉ signal-mediated activation of BDNF-PI and found that the Ca 2؉ -responsive elements were located in two separate (distal and proximal) regions and that the DNA sequences in the proximal region containing cAMP-responsive element (CRE), which is overlapped by the upstream stimulatory factor (USF)-binding element, were largely responsible for the activation of BDNF-PI. CRE-binding protein (CREB) family transcription factors and USF1/USF2 bind to this overlapping site, depending upon their preferred sequences which also control the magnitude of the activation. Overexpression of dominant negative CREB or USF reduced the BDNF-PI activation. These findings support that not only CREB but also USF1/USF2 contributes to Ca 2؉ signal-mediated activation of BDNF-PI through the recognition of an overlapping CRE and USF-binding element.

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Section: Calcium-responsive Elements Located In Two Distinctmentioning

confidence: 72%

Section: Methodsmentioning

confidence: 99%

Section: Production Of Bdnf Transcripts Containing Exons I Ii Iii mentioning

confidence: 99%

Section: Rna Isolation and Reverse Transcription (Rt)-pcr-totalmentioning

confidence: 99%

Section: Rna Isolation and Reverse Transcription (Rt)-pcr-totalmentioning

confidence: 99%

See 3 more Smart Citations

Involvement of an Upstream Stimulatory Factor as Well as cAMP-responsive Element-binding Protein in the Activation of Brain-derived Neurotrophic Factor Gene Promoter I

Tabuchi¹,

Sakaya²,

Kisukeda³

et al. 2002

Journal of Biological Chemistry

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197

172

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Identification of genetic networks involved in the cell growth arrest and differentiation of a rat astrocyte cell line RCG‐12

Takasaki

Takarada

Yasuda

et al. 2007

J of Cellular Biochemistry

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The purpose of the present study is to establish and characterize a conditionally immortalized astrocyte cell line and to clarify the genetic networks responsible for the cell growth arrest and differentiation. A conditionally immortalized astrocyte cell line, RCG-12, was established by infecting primary cultured rat cortical glia cells with a temperature-sensitive simian virus 40 large T-antigen. At a permissive temperature of 33 degrees C, the large T-antigen was expressed and cells grew continuously. On the other hand, the down-regulation of T-antigen at a non-permissive temperature of 39 degrees C led to growth arrest and differentiation. The cells expressed astrocyte-expressed genes such as glial fibrillary acidic protein. Interestingly, the differentiated condition induced by the non-permissive temperature significantly elevated the expression levels of several astrocyte-expressed genes. To identify the detailed mechanisms by which non-permissive temperature-induced cell growth arrest and differentiation, we performed high-density oligonucleotide microarray analysis and found that 556 out of 15,923 probe sets were differentially expressed 2.0-fold. A computational gene network analysis revealed that a genetic network containing up-regulated genes such as RB, NOTCH1, and CDKN1A was associated with the cellular growth and proliferation, and that a genetic network containing down-regulated genes such as MYC, CCNB1, and IGF1 was associated with the cell cycle. The established cell line RCG-12 retains some characteristics of astrocytes and should provide an excellent model for studies of astrocyte biology. The present results will also provide a basis for understanding the detailed molecular mechanisms of the growth arrest and differentiation of astrocytes.

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Silencer-Mediated Repression and Non-Mediated Activation of BDNF and c-fos Gene Promoters in Primary Glial or Neuronal Cells

Tabuchi

Nakatani

Nakaoka

et al. 1999

Biochemical and Biophysical Research Communications

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Inducibility of BDNF Gene Promoter I Detected by Calcium-Phosphate-Mediated DNA Transfection Is Confined to Neuronal but Not to Glial Cells

Cited by 29 publications

References 10 publications

Involvement of an Upstream Stimulatory Factor as Well as cAMP-responsive Element-binding Protein in the Activation of Brain-derived Neurotrophic Factor Gene Promoter I

Involvement of an Upstream Stimulatory Factor as Well as cAMP-responsive Element-binding Protein in the Activation of Brain-derived Neurotrophic Factor Gene Promoter I

Identification of genetic networks involved in the cell growth arrest and differentiation of a rat astrocyte cell line RCG‐12

Silencer-Mediated Repression and Non-Mediated Activation of BDNF and c-fos Gene Promoters in Primary Glial or Neuronal Cells

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