We examined intracellular pH (pH(i)) regulation in the retrotrapezoid nucleus (RTN), a CO(2)-sensitive site, and the hypoglossal nucleus, a nonchemosensitive site, during development (postnatal days 2-18) in rats. Respiratory acidosis [10% CO(2), extracellular pH (pH(o)) 7.18] caused acidification without pH(i) recovery in the RTN at all ages. In the hypoglossal nucleus, pH(i) recovered in young animals, but as animal age increased, the slope of pH(i) recovery diminished. In animals older than postnatal day 11, the pH(i) responses to hypercapnia were identical in the hypoglossal nucleus and the RTN, but hypoglossal nucleus and RTN neurons could regulate pH(i) during intracellular acidification at constant pH(o) at all ages. Recovery of pH(i) from acidification in the RTN depended on extracellular Na+ and was inhibited by amiloride but was unaffected by DIDS, suggesting a role for Na+/H+ exchange. Hence, pH(i) regulation during acidosis is more effective in the hypoglossal nucleus in younger animals, possibly as a requirement of development, but in older juvenile animals (older than postnatal day 11), pH(i) regulation is relatively poor in chemosensitive (RTN) and nonchemosensitive nuclei (hypoglossal nucleus).