Chemosensitivity of medullary neurons in explant tissue cultures

To determine whether cultured medulla contains chemosensitive neurons which are excited by CO2 and fixed acid and whether this function is specific to the ventral medulla, tissue explants of ventral and dorsal medulla were prepared from neonatal rats and incubated for two to three weeks. Cultures were superfused with artificial cerebrospinal fluid, maintained at 37 degrees C, and pH of the superfusate was varied either with PCO2 (14-71 Torr) at constant HCO3- (22 mM) or HCO3- (10-30 mM) at constant PCO2 (35 Torr). Spontaneous action potentials were recorded extracellularly in 51 ventral and 23 dorsal medullary neurons. Ventral medullary neurons exhibited a steady baseline firing frequency of 4 +/- 0.8 Hz. In contrast, dorsal medullary neurons exhibited two different patterns of spontaneous activity: 11 fired continuously (7.2 +/- 1.4 Hz) while 12 fired with a bursting pattern. Burst duration was 0.80 +/- 0.14 min and cycle time was 1.74 +/- 0.43 min. Decreasing pH with CO2 caused an increase in the activity of 10 of 27 ventral medullary neurons and two of six dorsal medullary neurons with a mean response of 7.5 Hz/pH unit. Varying pH by changing HCO3- had no effect on firing frequency. These results demonstrate that: (i) chemosensitive neurons are present in both ventral and dorsal medullary explant cultures; (ii) these cells only respond to changes in pH induced with CO2; and (iii) about half of the dorsal medullary neurons fire spontaneously with a regular bursting pattern of activity.