GFP-expressing locus ceruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture

Shereé M Johnson; Musa A Haxhiu; George B Richerson

doi:10.1152/japplphysiol.90414.2008

Back

GFP-expressing locus ceruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture

Journal article

Open access

Peer reviewed

GFP-expressing locus ceruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture

Shereé M Johnson, Musa A Haxhiu and George B Richerson

Journal of applied physiology (1985), Vol.105(4), pp.1301-1311

10/2008

DOI: 10.1152/japplphysiol.90414.2008

PMCID: PMC2576037

PMID: 18635881

Files and links (1)

url

https://doi.org/10.1152/japplphysiol.90414.2008View

Published (Version of record) Open Access

Abstract

The locus ceruleus (LC) contains neurons that increase their firing rate (FR) in vitro when exposed to elevated CO(2)/H(+) and have been proposed to influence the respiratory network to make compensatory adjustments in ventilation. Prp57 transgenic mice express green fluorescent protein (GFP) in the LC and were used to isolate, culture, and target LC neurons for electrophysiological recording. We hypothesized that GFP-LC neurons would exhibit CO(2)/H(+) chemosensitivity under primary culture conditions, evidenced as a change in FR. This is the first study to quantify CO(2)/H(+) responses in LC neuron FR in cell culture. Neurons were continuously bathed with solutions containing antagonists of glutamate and GABA receptors, and the acid-base status was changed from control (5% CO(2); pH approximately 7.4) to hypercapnic acidosis (9% CO(2); pH approximately 7.2) and hypocapnic alkalosis (3% CO(2); pH approximately 7.6). FR was quantified during perforated patch current clamp recordings. Approximately 86% of GFP-LC neurons were stimulated, and approximately 14% were insensitive to changes in CO(2)/H(+). The magnitude of the response of these neurons depended on the baseline FR, ranging from 155.9 +/- 6% when FR started at 2.95 +/- 0.49 Hz to 381 +/- 55.6% when FR started at 1.32 +/- 0.31 Hz. These results demonstrate that cultured LC neurons from Prp57 transgenic mice retain functional sensing molecules necessary for CO(2)/H(+) responses. Prp57 transgenic mice will serve as a valuable model to delineate mechanisms involved in CO(2)/H(+) responsiveness in catecholaminergic neurons.

Green Fluorescent Proteins - metabolism

Tyrosine 3-Monooxygenase - metabolism

Carbon Dioxide - metabolism

Catecholamines - metabolism

Chemoreceptor Cells - metabolism

Cells, Cultured

Mice, Transgenic

Green Fluorescent Proteins - genetics

Action Potentials

Patch-Clamp Techniques

Animals

Locus Coeruleus - cytology

Time Factors

Fluorescent Antibody Technique

Mice

Neurons - metabolism

Hydrogen-Ion Concentration

Locus Coeruleus - metabolism

Details

Title: Subtitle: GFP-expressing locus ceruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture
Creators: Shereé M Johnson - Department of Physiology and Biophysics, Howard University College of Medicine, 520 W Street Northwest, Washington, DC 20059, USA. smjohnson@howard.edu
Musa A Haxhiu
George B Richerson
Resource Type: Journal article
Publication Details: Journal of applied physiology (1985), Vol.105(4), pp.1301-1311
DOI: 10.1152/japplphysiol.90414.2008
PMID: 18635881
PMCID: PMC2576037
NLM abbreviation: J Appl Physiol (1985)
ISSN: 8750-7587
eISSN: 1522-1601
Publisher: United States
Grant note: NS 047422 / NINDS NIH HHS K01 NS047422-04 / NINDS NIH HHS NS 039407 / NINDS NIH HHS
Language: English
Date published: 10/2008
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Neurosurgery
Record Identifier: 9984020785602771

Metrics

19 Record Views

35 Times Cited - Web of Science