SummaryCO 2 is an important metabolic product whose concentrations are constantly monitored by CO 2 chemoreceptors. However, the high systemic CO 2 sensitivity may not be achieved by the CO 2 chemoreceptors without neuronal network processes. To show modulation of network properties during hypercapnia, we studied brainstem neurons dissociated from embryonic rats (P17-19) in multi-electrode arrays (MEA) after initial period (3 weeks) of culture. Spike trains of 33,622 pairs of units were analyzed using peri-event histograms (PEH). The amplitude of pericentral peaks between two CO 2 -stimulated units increased and the peak latency decreased during hypercapnia. Similar enhancement of synaptic strength was observed in those sharing a common input. These phenomena were not seen in CO 2 -unresponsive neurons. The amplitude of pericentral peaks between two CO 2 inhibited units also increased without changing latency. Over 60% CO 2 -stimulated neurons studied received mono-/oligosynaptic inputs from other CO 2 -stimulated cells, whereas only ~10% CO 2 -unresponsive neurons had such synaptic inputs. A small number of brainstem neurons showed electrical couplings. The coupling efficiency of CO 2 -stimulated but not CO 2 -unresponsive units was suppressed by ~50% with high PCO 2 . Inhibitory synaptic projections were also found, which was barely affected by hypercapnia. Consistent with the strengthening of excitatory synaptic connections, CO 2 sensitivity of post-synaptic neurons was significantly higher than presynaptic neurons. The difference was eliminated with blockade of presynaptic input. Based on these indirect assessments of synaptic interaction, our PEH analysis suggests that hypercapnia appears to modulate excitatory synaptic transmissions, especially those between CO 2 -stimulated neurons.
KeywordsCO 2 chemoreceptor; multielectrode arrays; MEA; peri-event histogram; cross-correlation; cell culture; neuronal network; electrical coupling
1, IntroductionCO 2 is a major metabolic product and plays an important role in systemic pH regulation and acid-base secretions. The PCO 2 levels are tightly regulated by several feedback control systems in which a critical step is the CO 2 detection by sensing cells in the brainstem and carotid body (Feldman et al. 2003;Richerson 2004;Putnam et al. 2004; Guyenet et al. 2005b). Of particular interest are brainstem CO 2 -chemosensitive neurons, as the systemic CO 2 response is retained after carotid bodies are removed bilaterally. With these CO 2 -chemosensitive cells, the nervous system can detect a change in arterial PCO 2 by as low as 1 torr and couple it to a 20-30% * Correspondence to: Phone: 404-651-0913, Fax: 404-651-2509, E-mail: cjiang@gsu.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Pl...