Differential effects of chronic drug treatment on alpha 3*and alpha 7 nicotinic receptor binding sites, in hippocampal neurones and SH-SY5Y cells

D L Ridley, A Rogers, S Wonnacott

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Abstract

1 The aim of this study was to compare the effects of chronic treatment (for 4 or 7 days) with nicotinic drugs and 20 mM KCl on numbers of surface alpha7 nicotinic AChR, identified by [I-125]-alpha bungarotoxin (alpha -Bgt) binding, in primary hippocampal cultures and SH-SY5Y cells. Numbers of alpha3* nicotinic AChR were also examined in SH-SY5Y cells, using [H-3]-epibatidine, which is predicted to label the total cellular population of predominantly alpha3 beta2* nicotinic AChR under the conditions used. 2 All the nicotinic agonists examined. the antagonists d-tubocurarine and methyllycaconitine, and KCl, upregulated[I-125]-alpha Bgt binding sites by 20-60% in hippocampal neurones and, where examined. SH-SY5Y cells. 3 Upregulation of [I-125]-alpha -Bgt binding sites by KCl was prevented by co-incubation with the L-type Ca2+ channel blocker verapamil or the Ca2+-calmodulin dependent kinase II (CaM-kinase II) inhibitor KN-62. Upregulation of [I-125]-alpha -Bgt binding sites by nicotine or 3,[(4-dimethylamino) cinnamylidene] anabaseine maleate (DMAC) was insensitive to these agents. 4 [H-3]-Epibatidine binding sites in SH-SY5Y cells were not affected by KCI but were upregulated in a verapamil-insensitive manner by nicotine and DMAC. KN-62 itself provoked a 2 fold increase in [H-3]-epibatidine binding. The inactive analogue KN-04 had no effect, suggesting that CaM-kinase II plays a role in regulating numbers of alpha3* nicotinic AChR. 5 These data indicate that numbers of alpha3* and alpha7 nicotinic AChR are modulated differently. Nicotinic agonists and KCI upregulate alpha7 nicotinic AChR through distinct cellular mechanisms, the latter involving L-type Ca2+ channels and CaM-kinase II. In contrast, alpha3* nicotinic AChR are not upregulated by KCI. This difference may reflect the distinct physiological roles proposed for alpha7 nicotinic AChR.
LanguageEnglish
Pages1286-1295
Number of pages10
JournalBritish Journal of Pharmacology
Volume133
Issue number8
DOIs
StatusPublished - 2001

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epibatidine
KN 62
Nicotinic Receptors
Bungarotoxins
Calcium-Calmodulin-Dependent Protein Kinases
Binding Sites
Nicotinic Agonists
Neurons
Up-Regulation
Verapamil
Nicotine
Pharmaceutical Preparations
Tubocurarine
Population

Cite this

@article{ab07e137a6b24c209ba5b6f4dc3adaa9,
title = "Differential effects of chronic drug treatment on alpha 3*and alpha 7 nicotinic receptor binding sites, in hippocampal neurones and SH-SY5Y cells",
abstract = "1 The aim of this study was to compare the effects of chronic treatment (for 4 or 7 days) with nicotinic drugs and 20 mM KCl on numbers of surface alpha7 nicotinic AChR, identified by [I-125]-alpha bungarotoxin (alpha -Bgt) binding, in primary hippocampal cultures and SH-SY5Y cells. Numbers of alpha3* nicotinic AChR were also examined in SH-SY5Y cells, using [H-3]-epibatidine, which is predicted to label the total cellular population of predominantly alpha3 beta2* nicotinic AChR under the conditions used. 2 All the nicotinic agonists examined. the antagonists d-tubocurarine and methyllycaconitine, and KCl, upregulated[I-125]-alpha Bgt binding sites by 20-60{\%} in hippocampal neurones and, where examined. SH-SY5Y cells. 3 Upregulation of [I-125]-alpha -Bgt binding sites by KCl was prevented by co-incubation with the L-type Ca2+ channel blocker verapamil or the Ca2+-calmodulin dependent kinase II (CaM-kinase II) inhibitor KN-62. Upregulation of [I-125]-alpha -Bgt binding sites by nicotine or 3,[(4-dimethylamino) cinnamylidene] anabaseine maleate (DMAC) was insensitive to these agents. 4 [H-3]-Epibatidine binding sites in SH-SY5Y cells were not affected by KCI but were upregulated in a verapamil-insensitive manner by nicotine and DMAC. KN-62 itself provoked a 2 fold increase in [H-3]-epibatidine binding. The inactive analogue KN-04 had no effect, suggesting that CaM-kinase II plays a role in regulating numbers of alpha3* nicotinic AChR. 5 These data indicate that numbers of alpha3* and alpha7 nicotinic AChR are modulated differently. Nicotinic agonists and KCI upregulate alpha7 nicotinic AChR through distinct cellular mechanisms, the latter involving L-type Ca2+ channels and CaM-kinase II. In contrast, alpha3* nicotinic AChR are not upregulated by KCI. This difference may reflect the distinct physiological roles proposed for alpha7 nicotinic AChR.",
author = "Ridley, {D L} and A Rogers and S Wonnacott",
year = "2001",
doi = "10.1038/sj.bjp.0704207",
language = "English",
volume = "133",
pages = "1286--1295",
journal = "British Journal of Pharmacology",
issn = "0007-1188",
publisher = "Wiley-Blackwell",
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TY - JOUR

T1 - Differential effects of chronic drug treatment on alpha 3*and alpha 7 nicotinic receptor binding sites, in hippocampal neurones and SH-SY5Y cells

AU - Ridley,D L

AU - Rogers,A

AU - Wonnacott,S

PY - 2001

Y1 - 2001

N2 - 1 The aim of this study was to compare the effects of chronic treatment (for 4 or 7 days) with nicotinic drugs and 20 mM KCl on numbers of surface alpha7 nicotinic AChR, identified by [I-125]-alpha bungarotoxin (alpha -Bgt) binding, in primary hippocampal cultures and SH-SY5Y cells. Numbers of alpha3* nicotinic AChR were also examined in SH-SY5Y cells, using [H-3]-epibatidine, which is predicted to label the total cellular population of predominantly alpha3 beta2* nicotinic AChR under the conditions used. 2 All the nicotinic agonists examined. the antagonists d-tubocurarine and methyllycaconitine, and KCl, upregulated[I-125]-alpha Bgt binding sites by 20-60% in hippocampal neurones and, where examined. SH-SY5Y cells. 3 Upregulation of [I-125]-alpha -Bgt binding sites by KCl was prevented by co-incubation with the L-type Ca2+ channel blocker verapamil or the Ca2+-calmodulin dependent kinase II (CaM-kinase II) inhibitor KN-62. Upregulation of [I-125]-alpha -Bgt binding sites by nicotine or 3,[(4-dimethylamino) cinnamylidene] anabaseine maleate (DMAC) was insensitive to these agents. 4 [H-3]-Epibatidine binding sites in SH-SY5Y cells were not affected by KCI but were upregulated in a verapamil-insensitive manner by nicotine and DMAC. KN-62 itself provoked a 2 fold increase in [H-3]-epibatidine binding. The inactive analogue KN-04 had no effect, suggesting that CaM-kinase II plays a role in regulating numbers of alpha3* nicotinic AChR. 5 These data indicate that numbers of alpha3* and alpha7 nicotinic AChR are modulated differently. Nicotinic agonists and KCI upregulate alpha7 nicotinic AChR through distinct cellular mechanisms, the latter involving L-type Ca2+ channels and CaM-kinase II. In contrast, alpha3* nicotinic AChR are not upregulated by KCI. This difference may reflect the distinct physiological roles proposed for alpha7 nicotinic AChR.

AB - 1 The aim of this study was to compare the effects of chronic treatment (for 4 or 7 days) with nicotinic drugs and 20 mM KCl on numbers of surface alpha7 nicotinic AChR, identified by [I-125]-alpha bungarotoxin (alpha -Bgt) binding, in primary hippocampal cultures and SH-SY5Y cells. Numbers of alpha3* nicotinic AChR were also examined in SH-SY5Y cells, using [H-3]-epibatidine, which is predicted to label the total cellular population of predominantly alpha3 beta2* nicotinic AChR under the conditions used. 2 All the nicotinic agonists examined. the antagonists d-tubocurarine and methyllycaconitine, and KCl, upregulated[I-125]-alpha Bgt binding sites by 20-60% in hippocampal neurones and, where examined. SH-SY5Y cells. 3 Upregulation of [I-125]-alpha -Bgt binding sites by KCl was prevented by co-incubation with the L-type Ca2+ channel blocker verapamil or the Ca2+-calmodulin dependent kinase II (CaM-kinase II) inhibitor KN-62. Upregulation of [I-125]-alpha -Bgt binding sites by nicotine or 3,[(4-dimethylamino) cinnamylidene] anabaseine maleate (DMAC) was insensitive to these agents. 4 [H-3]-Epibatidine binding sites in SH-SY5Y cells were not affected by KCI but were upregulated in a verapamil-insensitive manner by nicotine and DMAC. KN-62 itself provoked a 2 fold increase in [H-3]-epibatidine binding. The inactive analogue KN-04 had no effect, suggesting that CaM-kinase II plays a role in regulating numbers of alpha3* nicotinic AChR. 5 These data indicate that numbers of alpha3* and alpha7 nicotinic AChR are modulated differently. Nicotinic agonists and KCI upregulate alpha7 nicotinic AChR through distinct cellular mechanisms, the latter involving L-type Ca2+ channels and CaM-kinase II. In contrast, alpha3* nicotinic AChR are not upregulated by KCI. This difference may reflect the distinct physiological roles proposed for alpha7 nicotinic AChR.

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U2 - 10.1038/sj.bjp.0704207

DO - 10.1038/sj.bjp.0704207

M3 - Article

VL - 133

SP - 1286

EP - 1295

JO - British Journal of Pharmacology

T2 - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 8

ER -