Document Detail


GFP-expressing locus ceruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture.
MedLine Citation:
PMID:  18635881     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Shereé M Johnson; Musa A Haxhiu; George B Richerson
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2008-07-17
Journal Detail:
Title:  Journal of applied physiology (Bethesda, Md. : 1985)     Volume:  105     ISSN:  8750-7587     ISO Abbreviation:  J. Appl. Physiol.     Publication Date:  2008 Oct 
Date Detail:
Created Date:  2008-10-07     Completed Date:  2008-11-06     Revised Date:  2009-11-24    
Medline Journal Info:
Nlm Unique ID:  8502536     Medline TA:  J Appl Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1301-11     Citation Subset:  IM    
Affiliation:
Department of Physiology and Biophysics, Howard University College of Medicine, 520 W Street Northwest, Washington, DC 20059, USA. smjohnson@howard.edu
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MeSH Terms
Descriptor/Qualifier:
Action Potentials
Animals
Carbon Dioxide / metabolism*
Catecholamines / metabolism
Cells, Cultured
Chemoreceptor Cells / metabolism*
Fluorescent Antibody Technique
Green Fluorescent Proteins / genetics,  metabolism*
Hydrogen-Ion Concentration
Locus Coeruleus / cytology,  metabolism*
Mice
Mice, Transgenic
Neurons / metabolism*
Patch-Clamp Techniques
Time Factors
Tyrosine 3-Monooxygenase / metabolism
Grant Support
ID/Acronym/Agency:
K01 NS047422-04/NS/NINDS NIH HHS; NS 039407/NS/NINDS NIH HHS; NS 047422/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/Catecholamines; 124-38-9/Carbon Dioxide; 147336-22-9/Green Fluorescent Proteins; EC 1.14.16.2/Tyrosine 3-Monooxygenase
Comments/Corrections

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