Agonist-induced down-regulation of m1 muscarinic receptors and reduction of their mRNA level in a transfected cell line

Shou Zhen Wang; Jingru Hu; Rochelle M. Long; Wanida S. Pou; Carlos Forray; Esam E. El-Fakahany

doi:10.1016/0014-5793(90)80538-T

Agonist-induced down-regulation of m1 muscarinic receptors and reduction of their mRNA level in a transfected cell line

Shou Zhen Wang, Jingru Hu, Rochelle M. Long, Wanida S. Pou, Carlos Forray, Esam E. El-Fakahany

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Abstract

Agonist-induced reduction in both the number of ml muscarinic receptors and the mRNA coding for the receptor protein was investigated in Chinese hamster ovary (CHO) cells which were transfected with the ml muscarinic receptor gene. Receptor concentration was measured by the specific binding of the muscarinic ligand, [³H]quinuclidinyl benzilate ([³H]QNB), and Northern blot hybridization analysis was used to evaluate the levels of receptor mRNA. Incubation of cells with 1 mM of the muscarinic agonist, carbamylcholine (CBC), for 24 h decreased receptor density and mRNA levels in cells by 65% and 73%, respectively. These results indicate that agonist-induced down-regulation of ml muscarinic receptors might be due to, at least in part, a decrease in receptor synthesis resulting from a reduction in the steady-state level of their mRNA.

Original language	English (US)
Pages (from-to)	185-188
Number of pages	4
Journal	FEBS Letters
Volume	276
Issue number	1-2
DOIs	https://doi.org/10.1016/0014-5793(90)80538-T
State	Published - Dec 10 1990
Externally published	Yes

Bibliographical note

Funding Information:
AcknowledgementTs:h is work wass upportedin partb y NIH Grants NS-25743a ndA G-07118a ndby contractD AAL-03-88-0078fr om the US Army ResearchO ffice. E.E.E. is the recipiento f a Research Career DevelopmentA ward from NIH (AG-00344).T he authors would like to thankM r Mike Gentrya nd MS Donna Betheaf or their invaluablea ssistance.

Keywords

Chinese hamster ovary cell
Muscarinic receptor
Northern blot analysis
Receptor down-regulation
mRNA regulation

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title = "Agonist-induced down-regulation of m1 muscarinic receptors and reduction of their mRNA level in a transfected cell line",

abstract = "Agonist-induced reduction in both the number of ml muscarinic receptors and the mRNA coding for the receptor protein was investigated in Chinese hamster ovary (CHO) cells which were transfected with the ml muscarinic receptor gene. Receptor concentration was measured by the specific binding of the muscarinic ligand, [3H]quinuclidinyl benzilate ([3H]QNB), and Northern blot hybridization analysis was used to evaluate the levels of receptor mRNA. Incubation of cells with 1 mM of the muscarinic agonist, carbamylcholine (CBC), for 24 h decreased receptor density and mRNA levels in cells by 65% and 73%, respectively. These results indicate that agonist-induced down-regulation of ml muscarinic receptors might be due to, at least in part, a decrease in receptor synthesis resulting from a reduction in the steady-state level of their mRNA.",

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author = "Wang, {Shou Zhen} and Jingru Hu and Long, {Rochelle M.} and Pou, {Wanida S.} and Carlos Forray and El-Fakahany, {Esam E.}",

note = "Funding Information: AcknowledgementTs:h is work wass upportedin partb y NIH Grants NS-25743a ndA G-07118a ndby contractD AAL-03-88-0078fr om the US Army ResearchO ffice. E.E.E. is the recipiento f a Research Career DevelopmentA ward from NIH (AG-00344).T he authors would like to thankM r Mike Gentrya nd MS Donna Betheaf or their invaluablea ssistance.",

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doi = "10.1016/0014-5793(90)80538-T",

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AU - Wang, Shou Zhen

AU - Hu, Jingru

AU - Long, Rochelle M.

AU - Pou, Wanida S.

AU - Forray, Carlos

AU - El-Fakahany, Esam E.

N1 - Funding Information: AcknowledgementTs:h is work wass upportedin partb y NIH Grants NS-25743a ndA G-07118a ndby contractD AAL-03-88-0078fr om the US Army ResearchO ffice. E.E.E. is the recipiento f a Research Career DevelopmentA ward from NIH (AG-00344).T he authors would like to thankM r Mike Gentrya nd MS Donna Betheaf or their invaluablea ssistance.

PY - 1990/12/10

Y1 - 1990/12/10

N2 - Agonist-induced reduction in both the number of ml muscarinic receptors and the mRNA coding for the receptor protein was investigated in Chinese hamster ovary (CHO) cells which were transfected with the ml muscarinic receptor gene. Receptor concentration was measured by the specific binding of the muscarinic ligand, [3H]quinuclidinyl benzilate ([3H]QNB), and Northern blot hybridization analysis was used to evaluate the levels of receptor mRNA. Incubation of cells with 1 mM of the muscarinic agonist, carbamylcholine (CBC), for 24 h decreased receptor density and mRNA levels in cells by 65% and 73%, respectively. These results indicate that agonist-induced down-regulation of ml muscarinic receptors might be due to, at least in part, a decrease in receptor synthesis resulting from a reduction in the steady-state level of their mRNA.

AB - Agonist-induced reduction in both the number of ml muscarinic receptors and the mRNA coding for the receptor protein was investigated in Chinese hamster ovary (CHO) cells which were transfected with the ml muscarinic receptor gene. Receptor concentration was measured by the specific binding of the muscarinic ligand, [3H]quinuclidinyl benzilate ([3H]QNB), and Northern blot hybridization analysis was used to evaluate the levels of receptor mRNA. Incubation of cells with 1 mM of the muscarinic agonist, carbamylcholine (CBC), for 24 h decreased receptor density and mRNA levels in cells by 65% and 73%, respectively. These results indicate that agonist-induced down-regulation of ml muscarinic receptors might be due to, at least in part, a decrease in receptor synthesis resulting from a reduction in the steady-state level of their mRNA.

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KW - Receptor down-regulation

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Agonist-induced down-regulation of m1 muscarinic receptors and reduction of their mRNA level in a transfected cell line

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