title: Acidosis-stimulated neurons of the medullary raphe are serotonergic.
abstract: Neurons of the medullary raphe project widely to respiratory and autonomic nuclei and contain co-localized serotonin, thyrotropin-releasing hormone (TRH), and substance P, three neurotransmitters known to stimulate ventilation. Some medullary raphe neurons are highly sensitive to pH and CO(2) and have been proposed to be central chemoreceptors. Here it was determined whether these chemosensitive neurons are serotonergic. Cells were microdissected from the rat medullary raphe and maintained in primary cell culture for 13-70 days. Immunoreactivity for serotonin, substance P, and TRH was present in these cultures. All acidosis-stimulated neurons (n = 22) were immunoreactive for tryptophan hydroxylase (TpOH-IR), the rate-limiting enzyme for serotonin biosynthesis, whereas all acidosis-inhibited neurons (n = 16) were TpOH-immunonegative. The majority of TpOH-IR medullary raphe neurons (73%) were stimulated by acidosis. The electrophysiological properties of TpOH-IR neurons in culture were similar to those previously reported for serotonergic neurons in vivo and in brain slices. These properties included wide action potentials (4.55 +/- 0.5 ms) with a low variability of the interspike interval, a postspike afterhyperpolarization (AHP) that reversed 25 mV more positive than the Nernst potential for K(+), prominent A current, spike frequency adaptation and a prolonged AHP after a depolarizing pulse. Thus the intrinsic cellular properties of serotonergic neurons were preserved in cell culture, indicating that the results obtained using this in vitro approach are relevant to serotonergic neurons in vivo. These results demonstrate that acidosis-stimulated neurons of the medullary raphe contain serotonin. We propose that serotonergic neurons initiate a homeostatic response to changes in blood CO(2) that includes increased ventilation and modulation of autonomic function.
authors.affiliation: Departments of Neurology and Cellular and Molecular Physiology, Yale University, New Haven Connecticut, 06510, USA.
authors.forename: W
authors.initials: W
authors.lastname: Wang
authors.forename: J K
authors.initials: JK
authors.lastname: Tiwari
authors.forename: S R
authors.initials: SR
authors.lastname: Bradley
authors.forename: R V
authors.initials: RV
authors.lastname: Zaykin
authors.forename: G B
authors.initials: GB
authors.lastname: Richerson
mesh_terms: D000138:Acidosis / Q000097:blood / Q000503:physiopathology*; D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers; D002245:Carbon Dioxide / Q000097:blood; D002478:Cells, Cultured / Q000187:drug effects / Q000502:physiology; D002628:Chemoreceptor Cells / Q000187:drug effects / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000032:analysis; D009474:Neurons / Q000187:drug effects / Q000502:physiology; D010313:Partial Pressure; D018408:Patch-Clamp Techniques; D011498:Protein Precursors / Q000032:analysis; D011903:Raphe Nuclei / Q000187:drug effects* / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012143:Respiratory Physiological Phenomena; D012701:Serotonin / Q000032:analysis / Q000502:physiology*; D012890:Sleep / Q000502:physiology; D013373:Substance P / Q000032:analysis; D013973:Thyrotropin-Releasing Hormone / Q000032:analysis; D014365:Tryptophan Hydroxylase / Q000032:analysis
chemical_list: D015415:Biomarkers; D009419:Nerve Tissue Proteins; D011498:Protein Precursors; C063596:prepro-thyrotropin-releasing hormone; D002245:Carbon Dioxide; D012701:Serotonin; D013373:Substance P; D013973:Thyrotropin-Releasing Hormone; D014365:Tryptophan Hydroxylase