GFP-expressing locus ceruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture.
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.
Johnson
Shereé M
SM
Department of Physiology and Biophysics, Howard University College of Medicine, 520 W Street Northwest, Washington, DC 20059, USA. smjohnson@howard.edu
Haxhiu
Musa A
MA
Richerson
George B
GB
D000200:Action Potentials; D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism*; D002395:Catecholamines / Q000378:metabolism; D002478:Cells, Cultured; D002628:Chemoreceptor Cells / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008125:Locus Coeruleus / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D002395:Catecholamines; D002245:Carbon Dioxide; D049452:Green Fluorescent Proteins; D014446:Tyrosine 3-Monooxygenase