First Author | Coley AA | Year | 2013 |
Journal | Neuroscience | Volume | 246 |
Pages | 451-67 | PubMed ID | 23500099 |
Mgi Jnum | J:201360 | Mgi Id | MGI:5513060 |
Doi | 10.1016/j.neuroscience.2013.02.064 | Citation | Coley AA, et al. (2013) Immunocytochemical identification of electroneutral Na(+)-coupled HCO(3)(-) transporters in freshly dissociated mouse medullary raphe neurons. Neuroscience 246:451-67 |
abstractText | The medullary raphe (MR) of the medulla oblongata contains chemosensitive neurons that respond to increases in arterial [CO(2)], by altering firing rate, with increases being associated with serotonergic (5-hydroxytryptamine [5HT]) neurons and decreases, with GABAergic neurons. Both types of neurons contribute to increased alveolar ventilation. Decreases in intracellular pH are thought to link the rise in [CO(2)] to increased ventilation. Because electroneutral Na(+)-coupled HCO(3)(-) transporters (nNCBTs), which help protect cells from intracellular acidosis, are expressed robustly in the neurons of the central nervous system, a key question is whether these transporters are present in chemosensitive neurons. Therefore, we used an immunocytochemistry approach to identify neurons (using a microtubule associated protein-2 monoclonal antibody) and specifically 5HT neurons (TPH monoclonal antibody) or GABAergic neurons (GAD2 monoclonal antibody) in freshly dissociated cells from the mouse MR. We also co-labeled with polyclonal antibodies against the three nNCBTs: NBCn1, NDCBE, and NBCn2. We exploited ePet-EYFP (enhanced yellow fluorescent protein) mice (with EYFP-labeled 5HT neurons) as well as mice genetically deficient in each of the three nNCBTs. Quantitative image analysis distinguished positively stained cells from background signals. We found that >80% of GAD2(+) cells also were positive for NDCBE, and >90% of the TPH(+) and GAD2(+) cells were positive for the other nNCBTs. Assuming that the transporters are independently distributed among neurons, we can conclude that virtually all chemosensitive MR neurons contain at least one nNCBT. |