title: Regulation of ventral surface CO2/H+-sensitive neurons by purinergic signalling.
abstract: Central chemoreception is the mechanism by which the brain regulates breathing in response to changes in tissue CO(2)H+. A brainstem region called the retrotrapezoid nucleus (RTN) contains a population of CO2/H+-sensitive neurons that appears to function as an important chemoreceptor. Evidence also indicates that CO2-evoked ATP release from RTN astrocytes modulates activity of CO2/H+-sensitive neurons; however, the extent to which purinergic signalling contributes to chemoreception by RTN neurons is not clear and the mechanism(s) underlying CO2/H+-evoked ATP release is not fully elucidated. The goals of this study are to determine the extent to which ATP contributes to RTN chemoreception both in vivo and in vitro, and whether purinergic drive to chemoreceptors relies on extracellular Ca(2+) or gap junction hemichannels. We also examine the possible contribution of P2Y1 receptors expressed in the RTN to the purinergic drive to breathe.We show that purinergic signalling contributes, in part, to the CO(2)/H+ sensitivity of RTN neurons. In vivo, phrenic nerve recordings of respiratory activity in adult rats show that bilateral injections of pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS, a P2 receptor blocker) decreased the ventilatory response to CO2 by 30%. In vitro, loose-patch recordings from RTN neurons show that P2 receptor blockers decreased responsiveness to both 10% and 15% CO2 also by 30%. In the slice, the contribution of purinergic signalling to RTN chemoreception did not increase with temperature (22–35◦C) and was retained in low extracellular Ca2+ medium. Conversely, the gap junction blockers carbenoxolone and cobalt decreased neuronal CO2/H+ sensitivity by an amount similar to P2 receptor antagonists. Inhibition of the P2Y1 receptor in the RTN had no effect on CO2 responsivness in vitro or in vivo; thus, the identity of P2 receptors underlying the purinergic component of RTN chemoreception remains unknown. These results support the possibility that CO2/H+-evoked ATP release is mediated by a mechanism involving gap junction hemichannels.
authors.affiliation: University of Connecticut, Department of Physiology and Neurobiology, Storrs, CT 06269, USA. ian.wenker@uconn.edu
authors.forename: Ian C
authors.initials: IC
authors.lastname: Wenker
authors.forename: Cleyton R
authors.initials: CR
authors.lastname: Sobrinho
authors.forename: Ana C
authors.initials: AC
authors.lastname: Takakura
authors.forename: Thiago S
authors.initials: TS
authors.lastname: Moreira
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Mulkey
mesh_terms: D000200:Action Potentials; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001794:Blood Pressure; D001933:Brain Stem / Q000187:drug effects / Q000378:metabolism* / Q000503:physiopathology; D002118:Calcium / Q000378:metabolism; D002229:Carbenoxolone / Q000494:pharmacology; D002245:Carbon Dioxide / Q000378:metabolism*; D002628:Chemoreceptor Cells / Q000187:drug effects / Q000378:metabolism*; D003035:Cobalt / Q000494:pharmacology; D017630:Connexins / Q000378:metabolism; D003714:Denervation; D004195:Disease Models, Animal; D017629:Gap Junctions / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006935:Hypercapnia / Q000378:metabolism* / Q000503:physiopathology; D008297:Male; D018408:Patch-Clamp Techniques; D010791:Phrenic Nerve / Q000378:metabolism / Q000503:physiopathology; D058919:Purinergic P2 Receptor Antagonists / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D011983:Receptors, Purinergic / Q000187:drug effects / Q000378:metabolism*; D058472:Receptors, Purinergic P2Y1 / Q000378:metabolism; D012125:Respiratory Center / Q000378:metabolism / Q000503:physiopathology; D015656:Respiratory Mechanics; D015398:Signal Transduction / Q000187:drug effects; D013498:Suramin / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
chemical_list: D017630:Connexins; D058919:Purinergic P2 Receptor Antagonists; D011983:Receptors, Purinergic; D058472:Receptors, Purinergic P2Y1; D002245:Carbon Dioxide; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D003035:Cobalt; D011732:Pyridoxal Phosphate; D013498:Suramin; D000255:Adenosine Triphosphate; D002229:Carbenoxolone; D002118:Calcium