Binding of [3H]dihydroalprenolol and [3H]quinuclidinyl benzilate to intact cells of cultured corneal epithelium

A. M. Colley, Harrison D Cavanagh

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Abstract

In intact cultured rabbit corneal cells we have identified [3H]dihydroalprenolol ([3H]DHA) and [3H]quinuclidinyl benzilate ([3H]QNB) binding activities which meet criteria for β-adrenergic and muscarinic cholinergic receptors. For saturable, propranolol-sensitive [3H]DHA binding, B(max)=0.374 ± 0.063 fmol/μg protein; K(DHA)=12.5 ± 2.4 nM from Scatchard analysis. For saturable, atropine-sensitive [3H]QNB binding, B(max)=0.403 ± 0.053 fmol/μg protein; K(QNB)=15.4 ± 0.7 nM. The order of potency of unlabeled adrenergic agonists in competition for [3H]DHA sites was isoproterenol>epinephrine>norepinephrine. For unlabeled cholinergic agonists competing for [3H]QNB sites, the order was oxotremorine>acetylcholine≥carbamylcholine. Acetylcholine did not inhibit [3H]DHA binding, nor did isoproterenol or choline inhibit [3H]QNB binding. Effectiveness of drugs in stimulating cAMP or cGMP accumulation closely paralleled efficacy in competition for [3H]DHA or [3H]QNB sites. Results confirm the presence in intact cultured corneal membrane suspensions), identify in intact cells muscarinic cholinergic receptors (not previously detected in broken cell preparations), and supply evidence for receptor-mediated regulation of cyclic nucleotide levels in these cells, further supporting our hypothesis of bidirectional influence by cAMP-mediated β-adrenergic and cGMP-mediated cholinergic 'first messengers' on proliferation during healing of corneal epithelial defects.

Original languageEnglish (US)
Pages (from-to)75-86
Number of pages12
JournalMetabolic, Pediatric and Systemic Ophthalmology
Volume6
Issue number2
StatePublished - 1982

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Dihydroalprenolol
Quinuclidinyl Benzilate
Corneal Epithelium
Cultured Cells
Cholinergic Receptors
Muscarinic Receptors
Isoproterenol
Adrenergic Agents
Oxotremorine
Cholinergic Agonists
Adrenergic Agonists
Cyclic Nucleotides
Choline
Atropine
Propranolol
Cholinergic Agents
Epinephrine
Acetylcholine
Suspensions
Norepinephrine

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Ophthalmology
  • Pediatrics, Perinatology, and Child Health

Cite this

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title = "Binding of [3H]dihydroalprenolol and [3H]quinuclidinyl benzilate to intact cells of cultured corneal epithelium",
abstract = "In intact cultured rabbit corneal cells we have identified [3H]dihydroalprenolol ([3H]DHA) and [3H]quinuclidinyl benzilate ([3H]QNB) binding activities which meet criteria for β-adrenergic and muscarinic cholinergic receptors. For saturable, propranolol-sensitive [3H]DHA binding, B(max)=0.374 ± 0.063 fmol/μg protein; K(DHA)=12.5 ± 2.4 nM from Scatchard analysis. For saturable, atropine-sensitive [3H]QNB binding, B(max)=0.403 ± 0.053 fmol/μg protein; K(QNB)=15.4 ± 0.7 nM. The order of potency of unlabeled adrenergic agonists in competition for [3H]DHA sites was isoproterenol>epinephrine>norepinephrine. For unlabeled cholinergic agonists competing for [3H]QNB sites, the order was oxotremorine>acetylcholine≥carbamylcholine. Acetylcholine did not inhibit [3H]DHA binding, nor did isoproterenol or choline inhibit [3H]QNB binding. Effectiveness of drugs in stimulating cAMP or cGMP accumulation closely paralleled efficacy in competition for [3H]DHA or [3H]QNB sites. Results confirm the presence in intact cultured corneal membrane suspensions), identify in intact cells muscarinic cholinergic receptors (not previously detected in broken cell preparations), and supply evidence for receptor-mediated regulation of cyclic nucleotide levels in these cells, further supporting our hypothesis of bidirectional influence by cAMP-mediated β-adrenergic and cGMP-mediated cholinergic 'first messengers' on proliferation during healing of corneal epithelial defects.",
author = "Colley, {A. M.} and Cavanagh, {Harrison D}",
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AU - Colley, A. M.

AU - Cavanagh, Harrison D

PY - 1982

Y1 - 1982

N2 - In intact cultured rabbit corneal cells we have identified [3H]dihydroalprenolol ([3H]DHA) and [3H]quinuclidinyl benzilate ([3H]QNB) binding activities which meet criteria for β-adrenergic and muscarinic cholinergic receptors. For saturable, propranolol-sensitive [3H]DHA binding, B(max)=0.374 ± 0.063 fmol/μg protein; K(DHA)=12.5 ± 2.4 nM from Scatchard analysis. For saturable, atropine-sensitive [3H]QNB binding, B(max)=0.403 ± 0.053 fmol/μg protein; K(QNB)=15.4 ± 0.7 nM. The order of potency of unlabeled adrenergic agonists in competition for [3H]DHA sites was isoproterenol>epinephrine>norepinephrine. For unlabeled cholinergic agonists competing for [3H]QNB sites, the order was oxotremorine>acetylcholine≥carbamylcholine. Acetylcholine did not inhibit [3H]DHA binding, nor did isoproterenol or choline inhibit [3H]QNB binding. Effectiveness of drugs in stimulating cAMP or cGMP accumulation closely paralleled efficacy in competition for [3H]DHA or [3H]QNB sites. Results confirm the presence in intact cultured corneal membrane suspensions), identify in intact cells muscarinic cholinergic receptors (not previously detected in broken cell preparations), and supply evidence for receptor-mediated regulation of cyclic nucleotide levels in these cells, further supporting our hypothesis of bidirectional influence by cAMP-mediated β-adrenergic and cGMP-mediated cholinergic 'first messengers' on proliferation during healing of corneal epithelial defects.

AB - In intact cultured rabbit corneal cells we have identified [3H]dihydroalprenolol ([3H]DHA) and [3H]quinuclidinyl benzilate ([3H]QNB) binding activities which meet criteria for β-adrenergic and muscarinic cholinergic receptors. For saturable, propranolol-sensitive [3H]DHA binding, B(max)=0.374 ± 0.063 fmol/μg protein; K(DHA)=12.5 ± 2.4 nM from Scatchard analysis. For saturable, atropine-sensitive [3H]QNB binding, B(max)=0.403 ± 0.053 fmol/μg protein; K(QNB)=15.4 ± 0.7 nM. The order of potency of unlabeled adrenergic agonists in competition for [3H]DHA sites was isoproterenol>epinephrine>norepinephrine. For unlabeled cholinergic agonists competing for [3H]QNB sites, the order was oxotremorine>acetylcholine≥carbamylcholine. Acetylcholine did not inhibit [3H]DHA binding, nor did isoproterenol or choline inhibit [3H]QNB binding. Effectiveness of drugs in stimulating cAMP or cGMP accumulation closely paralleled efficacy in competition for [3H]DHA or [3H]QNB sites. Results confirm the presence in intact cultured corneal membrane suspensions), identify in intact cells muscarinic cholinergic receptors (not previously detected in broken cell preparations), and supply evidence for receptor-mediated regulation of cyclic nucleotide levels in these cells, further supporting our hypothesis of bidirectional influence by cAMP-mediated β-adrenergic and cGMP-mediated cholinergic 'first messengers' on proliferation during healing of corneal epithelial defects.

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