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J Neurophysiol 85: 2224-2235, 2001;
0022-3077/01 $5.00
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The Journal of Neurophysiology Vol. 85 No. 5 May 2001, pp. 2224-2235
Copyright ©2001 by the American Physiological Society

Acidosis-Stimulated Neurons of the Medullary Raphe Are Serotonergic

Wengang Wang,1 Jyoti K. Tiwari,1 Stefania Risso Bradley,1 Rey V. Zaykin,1 and George B. Richerson1,2

 1Departments of Neurology and Cellular and Molecular Physiology, Yale University, New Haven 06510; and  2Veteran's Affairs Medical Center, West Haven, Connecticut 06516

Wang, Wengang, Jyoti K. Tiwari, Stefania Risso Bradley, Rey V. Zaykin, and George B. Richerson. Acidosis-Stimulated Neurons of the Medullary Raphe Are Serotonergic. J. Neurophysiol. 85: 2224-2235, 2001. 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 CO2 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 CO2 that includes increased ventilation and modulation of autonomic function.




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