Muscarinic receptors and second-messenger responses of neurons in primary culture

Ellis, John E.; Huyler, Joanne H.; Kemp, Dorothy E.; Weiss, Samuel

doi:10.1016/0006-8993(90)90167-a

Cited by 18 publications

(7 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These resemble published values for brain tissues (e.g. Rooney and Nahorski, 1986), cultured neurons (Gonzales et al, 1985;Ellis et a/., 1990) and cells transfected with mAChR subtypes (e.g. Peralta et al, 1987).…”

Section: Rnachr-mediated Phosphoinositide Turnoversupporting

confidence: 84%

“…Published studies (e.g. Forray and El-Fakahany, 1990;Ellis et al, 1990;McKinney et al, 1991) have shown that this makes it possible to obtain good estimates of the antagonist affinities in inositol phosphate responses in brain tissues since the mAChRs coupled to the phosphoinositide metabolism in most brain regions have small receptor reserves (Rooney and Nahorski, 1986;Fisher and Snider, 1987). 250 pg/ml denatured salmon sperm DNA per ml for -18 h, hybridized in the same medium with the ["PI probes (-1 X lo6 c.p.m./ml) for at least 16 h, then washed twice for I h with IXSSCI5XDenhardt's solution/0.5% SDS and finally once with 0.2XSSC/0.5% SDS for 30 min.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Cultured Neurons from Mouse Brain Reproduce the Muscarinic Receptor Profile of Their Tissue of Origin

André

Santos

Koulakoff

1994

Eur J of Neuroscience

View full text Add to dashboard Cite

These studies investigate the regional variations in the muscarinic acetylcholine receptor (mAChR) profiles in neuron populations of the CNS using primary neuron cultures derived from three areas of the mouse brain--the cerebral hemispheres, the mesencephalon and the medulla-pons--that have distinct mAChR systems. We first assessed the extent to which neurons reproduced their in vivo properties in culture by monitoring the binding capacity, the pharmacological profiles and the levels of mAChR transcripts in neuron cultures and their tissues of origin. We showed that the primary neuron cultures accumulated mAChRs with initial rates similar to those in vivo, had pharmacological profiles very close to those of their area of origin, and accumulated m1, m2, m3, m4 and m5 receptor transcripts according to patterns resembling those in the tissues. We conclude that most of the characteristics of the mAChRs in a given area are proper to the neuron population of that area, that the pattern is established early in ontogenesis, and that it is reproduced in vitro. We also show that the stimulation of phosphoinositide turnover is mediated by mAChRs with distinct pharmacological profiles in neuron cultures from the three brain areas.

show abstract

Section: Rnachr-mediated Phosphoinositide Turnoversupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Cultured Neurons from Mouse Brain Reproduce the Muscarinic Receptor Profile of Their Tissue of Origin

André

Santos

Koulakoff

1994

Eur J of Neuroscience

View full text Add to dashboard Cite

show abstract

“…The protocol used for measuring inositol phosphate (IP) metabolism was adapted from Ellis et al (1990) and Bymaster et al (1999). In brief, cells were cultured in 24-well plates (Falcon; BD Biosciences Discovery Labware, Bedford, MA) at a density of 5 ϫ10 4 cells/well in 0.5 ml of F-12 medium.…”

Section: Methodsmentioning

confidence: 99%

Allosteric Modulation of the M₃ Muscarinic Receptor by Amiodarone and N-Ethylamiodarone: Application of the Four-Ligand Allosteric Two-State Model

Stahl¹,

Elmslie²,

Ellis³

2011

Mol Pharmacol

Self Cite

View full text Add to dashboard Cite

We have reported previously that amiodarone interacts with muscarinic receptors via a novel allosteric site. This study presents mechanistic details on the nature of that interaction. Amiodarone enhanced the maximal level of agonist-stimulated release of arachidonic acid (AA) from Chinese hamster ovary cells that expressed M 3 muscarinic receptors; this enhancement was observed for acetylcholine and for the partial agonist pilocarpine. A similar effect of amiodarone was observed when pilocarpine was used to stimulate inositol phosphate (IP) metabolism, but not when acetylcholine was used. Subsequent studies showed that the IP response exhibited a much larger receptor reserve than the AA response, and reduction of that reserve by receptor alkylation unmasked amiodarone's enhancement of the maximal IP response to acetylcholine. Modulating the receptor reserve also revealed acetylcholine's greater affinity (K A ) for the conformation of the receptor that mediates the AA response. The amiodarone analog N-ethylamiodarone (NEA) did not alter maximal agonist response but merely reduced agonist potency (that is, it appeared to be an antagonist). However, the action of NEA could be clearly distinguished from the action of the orthosteric antagonist NMS. Demonstration of this point was facilitated by an elaboration of Hall's allosteric two-state model; this new model represents a system composed of two ligands that compete with each other at the orthosteric site and two ligands that compete with each other at the allosteric site. In conclusion, amiodarone competes with NEA at a novel, extracellular, allosteric site to enhance the maximal stimulation evoked by acetylcholine and pilocarpine in two different responses.

show abstract

“…Published studies (e.g. Ellis et al, 1990;Forray and El-Fakahany, 1990;McKinney et al, 1991) have shown that this correction gives good estimates of the antagonist affinities in the inositol phosphate responses in brain tissues since the mAChRs coupled to the phosphoinositide metabolism in most brain regions show only a small amount of receptor reserve (Rooney and Nahorski, 1986;Fisher and Snider, 1987).…”

Section: Data Anulysismentioning

confidence: 99%

Muscarinic Receptor Profiles of Mouse Brain Astrocytes in Culture Vary With Their Tissue Of Origin but Differ From Those of Neurons

André

Santos

Koulakoff

1994

Eur J of Neuroscience

View full text Add to dashboard Cite

The two main cell populations in brain tissues are neurons and astrocytes. Cultures of both bear muscarinic acetylcholine receptors (mAChRs). Available data indicate that astrocyte mACRs are heterogeneous, but the particular subtypes on these cells are not known, nor is there any information as to whether there is a regional variation in the mAChR profile of astrocytes. This paper describes the mAChR profiles of cultured astrocytes derived from the cerebral hemispheres, mesencephalon and medulla-pons, and is a continuation of our study on cultures of neurons from these same tissues. Pharmacological studies showed that astrocytes accumulated small amounts of mAChRs with distinct pharmacological profiles which, for a given area, differed from those of neurons in culture. Northern blot analyses showed transcripts for m1 and m3 mAChRs only. Their concentrations differed from one cell population to another. Astrocyte cultures from the mesencephalon contained m1 mRNA amounts close to those in the tissue. Thus, at least part of the mAChR profile in vitro might be a true reflection of the cell's properties in vivo. Functional studies showed that mAChRs mediate the stimulation of phosphoinositide turnover in all three astrocyte cultures, that the amplitude of this response varies greatly with the origin of the cell, and that two pharmacological subclasses, M1 and M1-2-, are involved in these responses, but to different extents. Thus the CNS contains discrete astrocyte populations which in culture differ in their mAChR profiles at the molecular, the pharmacological and the functional levels.

show abstract

Muscarinic receptors and second-messenger responses of neurons in primary culture

Cited by 18 publications

References 25 publications

Cultured Neurons from Mouse Brain Reproduce the Muscarinic Receptor Profile of Their Tissue of Origin

Cultured Neurons from Mouse Brain Reproduce the Muscarinic Receptor Profile of Their Tissue of Origin

Allosteric Modulation of the M₃ Muscarinic Receptor by Amiodarone and N-Ethylamiodarone: Application of the Four-Ligand Allosteric Two-State Model

Muscarinic Receptor Profiles of Mouse Brain Astrocytes in Culture Vary With Their Tissue Of Origin but Differ From Those of Neurons

Contact Info

Product

Resources

About

Muscarinic receptors and second-messenger responses of neurons in primary culture

Cited by 18 publications

References 25 publications

Cultured Neurons from Mouse Brain Reproduce the Muscarinic Receptor Profile of Their Tissue of Origin

Cultured Neurons from Mouse Brain Reproduce the Muscarinic Receptor Profile of Their Tissue of Origin

Allosteric Modulation of the M3 Muscarinic Receptor by Amiodarone and N-Ethylamiodarone: Application of the Four-Ligand Allosteric Two-State Model

Muscarinic Receptor Profiles of Mouse Brain Astrocytes in Culture Vary With Their Tissue Of Origin but Differ From Those of Neurons

Contact Info

Product

Resources

About

Allosteric Modulation of the M₃ Muscarinic Receptor by Amiodarone and N-Ethylamiodarone: Application of the Four-Ligand Allosteric Two-State Model