Postnatal development of N-methyl- d-aspartate receptor subunits 2A, 2B, 2C, 2D, and 3B immunoreactivity in brain stem respiratory nuclei of the rat

Abstract

Previously, we reported that a critical period in respiratory network development exists in rats around postnatal days (P; P12–P13), when abrupt neurochemical, metabolic, and physiological changes occur. Specifically, the expressions of glutamate and N-methyl- d-aspartate (NMDA) receptor (NR) subunit 1 in the pre-Bötzinger complex (PBC), nucleus ambiguus (Amb), hypoglossal nucleus (XII), and ventrolateral subnucleus of solitary tract nucleus (NTS VL) were significantly reduced at P12. To test our hypothesis that other NR subunits also undergo postnatal changes, we undertook an in-depth immunohistochemical study of NR2A, 2B, 2C, 2D, and 3B in these four respiratory nuclei in P2–P21 rats, using the non-respiratory cuneate nucleus (CN) as a control. Our results revealed that: (1) NR2A expression increased gradually from P2 to P11, but fell significantly at P12 in all four respiratory nuclei (but not in the CN), followed by a quick rise and a relative plateau until P21; (2) NR2B expression remained relatively constant from P2 to P21 in all five nuclei examined; (3) NR2C expression had an initial rise from P2 to P3, but remained relatively constant thereafter until P21, except for a significant fall at P12 in the PBC; (4) NR2D expression fell significantly from P2 to P3, then plateaued until P12, and declined again until P21; and (5) in contrast to NR2D, NR3B expression rose gradually from P2 to P21. These patterns reflect a dynamic remodeling of NMDA receptor subunit composition during postnatal development, with a distinct reduction of NR2A expression during the critical period (P12), just as NR1 did in various respiratory nuclei. There was also a potential switch between the neonatal NR2D and the more mature NR3B subunit, possibly around the critical period. Thus, during the critical period, NMDA receptors are undergoing greater adjustments that may contribute to attenuated excitatory synaptic transmission in the respiratory network.