Differential promoter activity of the mouse apolipoprotein (APOE) gene in primary neurons, astrocytoma and in PC12 cells

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Abstract

The APOE gene, which constitutes a major susceptibility factor for the development of the familial and sporadic forms of late-onset Alzheimer's disease (AD), encodes a 34 kDa protein. It plays a critical role in mobilization and redistribution of cholesterol and phospholipid during membrane remodeling and synaptic plasticity. The gene is located on 19q13 of the human chromosome. The proximal 5′-flanking region of the APOE gene is highly conserved in the mouse, rat and human; the relative position of the TATA box' and the two copies of 'GC box' are identical. To study the transcription control of the mouse (m) APOE gene, we assayed in different cell types the promoter activity of a 725 nucleotide (nt) 5′-flanking region, which is located 772 nt upstream from the translation initiation codon. We cloned the 725 nt region into a promoterless vector upstream of the reporter chloramphenicol acetyl transferase (CAT) gene. The mAPOE promoter and vector DNAs were independently transfected in primary rat cortical neurons, human astrocytoma(U138) and PC 12 cell lines. In mAPOE-transfected U-138 cells, we observed a 5-fold increase in CAT reporter activity from the promoterless vector. As compared to U138 cells, we detected a reduced CAT activity in rat cortical neurons and PC 12 cell lines. In both these cells, the mAPOE promoter displayed significantly higher levels of activity than the vector. Our results suggest that mAPOE can also be expressed in neuronal cells in addition to the astrocytic cells. Characterization of mAPOE promoter is important for the APOE transgenic mice studies, which are used for the AD drug development discovery.

Original languageEnglish
Pages (from-to)589-590
Number of pages2
JournalAmerican Journal of Medical Genetics, Part B: Neuropsychiatric Genetics
Volume105
Issue number7
StatePublished - Oct 8 2001

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Apolipoproteins
PC12 Cells
Astrocytoma
Neurons
Chloramphenicol
Transferases
Genes
Nucleotides
5' Flanking Region
Alzheimer Disease
Cell Line
TATA Box
Neuronal Plasticity
Initiator Codon
Human Chromosomes
Drug Discovery
Transgenic Mice
Phospholipids
Cholesterol
Membranes

ASJC Scopus subject areas

  • Genetics(clinical)
  • Neuropsychology and Physiological Psychology
  • Neuroscience(all)

Cite this

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title = "Differential promoter activity of the mouse apolipoprotein (APOE) gene in primary neurons, astrocytoma and in PC12 cells",
abstract = "The APOE gene, which constitutes a major susceptibility factor for the development of the familial and sporadic forms of late-onset Alzheimer's disease (AD), encodes a 34 kDa protein. It plays a critical role in mobilization and redistribution of cholesterol and phospholipid during membrane remodeling and synaptic plasticity. The gene is located on 19q13 of the human chromosome. The proximal 5′-flanking region of the APOE gene is highly conserved in the mouse, rat and human; the relative position of the TATA box' and the two copies of 'GC box' are identical. To study the transcription control of the mouse (m) APOE gene, we assayed in different cell types the promoter activity of a 725 nucleotide (nt) 5′-flanking region, which is located 772 nt upstream from the translation initiation codon. We cloned the 725 nt region into a promoterless vector upstream of the reporter chloramphenicol acetyl transferase (CAT) gene. The mAPOE promoter and vector DNAs were independently transfected in primary rat cortical neurons, human astrocytoma(U138) and PC 12 cell lines. In mAPOE-transfected U-138 cells, we observed a 5-fold increase in CAT reporter activity from the promoterless vector. As compared to U138 cells, we detected a reduced CAT activity in rat cortical neurons and PC 12 cell lines. In both these cells, the mAPOE promoter displayed significantly higher levels of activity than the vector. Our results suggest that mAPOE can also be expressed in neuronal cells in addition to the astrocytic cells. Characterization of mAPOE promoter is important for the APOE transgenic mice studies, which are used for the AD drug development discovery.",
author = "Debomoy Lahiri and Ge, {Y. W.} and X. Chen and John Nurnberger and Martin Farlow and Yansheng Du",
year = "2001",
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T1 - Differential promoter activity of the mouse apolipoprotein (APOE) gene in primary neurons, astrocytoma and in PC12 cells

AU - Lahiri, Debomoy

AU - Ge, Y. W.

AU - Chen, X.

AU - Nurnberger, John

AU - Farlow, Martin

AU - Du, Yansheng

PY - 2001/10/8

Y1 - 2001/10/8

N2 - The APOE gene, which constitutes a major susceptibility factor for the development of the familial and sporadic forms of late-onset Alzheimer's disease (AD), encodes a 34 kDa protein. It plays a critical role in mobilization and redistribution of cholesterol and phospholipid during membrane remodeling and synaptic plasticity. The gene is located on 19q13 of the human chromosome. The proximal 5′-flanking region of the APOE gene is highly conserved in the mouse, rat and human; the relative position of the TATA box' and the two copies of 'GC box' are identical. To study the transcription control of the mouse (m) APOE gene, we assayed in different cell types the promoter activity of a 725 nucleotide (nt) 5′-flanking region, which is located 772 nt upstream from the translation initiation codon. We cloned the 725 nt region into a promoterless vector upstream of the reporter chloramphenicol acetyl transferase (CAT) gene. The mAPOE promoter and vector DNAs were independently transfected in primary rat cortical neurons, human astrocytoma(U138) and PC 12 cell lines. In mAPOE-transfected U-138 cells, we observed a 5-fold increase in CAT reporter activity from the promoterless vector. As compared to U138 cells, we detected a reduced CAT activity in rat cortical neurons and PC 12 cell lines. In both these cells, the mAPOE promoter displayed significantly higher levels of activity than the vector. Our results suggest that mAPOE can also be expressed in neuronal cells in addition to the astrocytic cells. Characterization of mAPOE promoter is important for the APOE transgenic mice studies, which are used for the AD drug development discovery.

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