Molecular cloning of a novel protein regulated by opioid treatment of NG108-15 cells

Marilee J. Wick; David K. Ann; Horace H Loh

doi:10.1016/0169-328X(95)00090-F

Molecular cloning of a novel protein regulated by opioid treatment of NG108-15 cells

Marilee J. Wick, David K. Ann, Horace H Loh

Pharmacology

Research output: Contribution to journal › Article › peer-review

20 Scopus citations

Abstract

A new cDNA clone, NGD5, has been identified from a subtraction cDNA library constructed with mRNA isolated from control neuroblastoma X glioma NG108-15 cells and cells treated for 48 h with the δ-opioid agonist, d-Ala², d-Leu⁵ enkephalin (DADLE). NGD5 mRNA is decreased, in a naloxone-reversible manner, upon long-term treatment of NG108-15 cells with DADLE, indicating that this clone may be related to opioid receptor function. Northern analysis indicates that NGD5 mRNA is expressed in rat brain. Two similar nearly full-length NGD5 clones, NGD5A and NGD5B, were isolated from a λgt10 NG108-15 cDNA library and sequenced. The predicted 40-kDa peptide encoded for by the NGD5 cDNA has no significant homology to the recently cloned μ δ or κ opioid receptors nor to any other known proteins.

Original language	English (US)
Pages (from-to)	171-175
Number of pages	5
Journal	Molecular Brain Research
Volume	32
Issue number	1
DOIs	https://doi.org/10.1016/0169-328X(95)00090-F
State	Published - Aug 1995

Keywords

Down regulation
Neuroblastoma cell
Subtractive cDNA cloning
δ-Opioid receptor

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title = "Molecular cloning of a novel protein regulated by opioid treatment of NG108-15 cells",

abstract = "A new cDNA clone, NGD5, has been identified from a subtraction cDNA library constructed with mRNA isolated from control neuroblastoma X glioma NG108-15 cells and cells treated for 48 h with the δ-opioid agonist, d-Ala2, d-Leu5 enkephalin (DADLE). NGD5 mRNA is decreased, in a naloxone-reversible manner, upon long-term treatment of NG108-15 cells with DADLE, indicating that this clone may be related to opioid receptor function. Northern analysis indicates that NGD5 mRNA is expressed in rat brain. Two similar nearly full-length NGD5 clones, NGD5A and NGD5B, were isolated from a λgt10 NG108-15 cDNA library and sequenced. The predicted 40-kDa peptide encoded for by the NGD5 cDNA has no significant homology to the recently cloned μ δ or κ opioid receptors nor to any other known proteins.",

keywords = "Down regulation, Neuroblastoma cell, Subtractive cDNA cloning, δ-Opioid receptor",

author = "Wick, {Marilee J.} and Ann, {David K.} and Loh, {Horace H}",

year = "1995",

month = aug,

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AU - Wick, Marilee J.

AU - Ann, David K.

AU - Loh, Horace H

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N2 - A new cDNA clone, NGD5, has been identified from a subtraction cDNA library constructed with mRNA isolated from control neuroblastoma X glioma NG108-15 cells and cells treated for 48 h with the δ-opioid agonist, d-Ala2, d-Leu5 enkephalin (DADLE). NGD5 mRNA is decreased, in a naloxone-reversible manner, upon long-term treatment of NG108-15 cells with DADLE, indicating that this clone may be related to opioid receptor function. Northern analysis indicates that NGD5 mRNA is expressed in rat brain. Two similar nearly full-length NGD5 clones, NGD5A and NGD5B, were isolated from a λgt10 NG108-15 cDNA library and sequenced. The predicted 40-kDa peptide encoded for by the NGD5 cDNA has no significant homology to the recently cloned μ δ or κ opioid receptors nor to any other known proteins.

AB - A new cDNA clone, NGD5, has been identified from a subtraction cDNA library constructed with mRNA isolated from control neuroblastoma X glioma NG108-15 cells and cells treated for 48 h with the δ-opioid agonist, d-Ala2, d-Leu5 enkephalin (DADLE). NGD5 mRNA is decreased, in a naloxone-reversible manner, upon long-term treatment of NG108-15 cells with DADLE, indicating that this clone may be related to opioid receptor function. Northern analysis indicates that NGD5 mRNA is expressed in rat brain. Two similar nearly full-length NGD5 clones, NGD5A and NGD5B, were isolated from a λgt10 NG108-15 cDNA library and sequenced. The predicted 40-kDa peptide encoded for by the NGD5 cDNA has no significant homology to the recently cloned μ δ or κ opioid receptors nor to any other known proteins.

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KW - Neuroblastoma cell

KW - Subtractive cDNA cloning

KW - δ-Opioid receptor

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