Expression of N-methyl-D-aspartate receptor and its effect on nitric oxide production of rat alveolar macrophages
Related Experiment Video
Updated: Jun 8, 2026
Isolation and In vitro Culture of Bone Marrow-Derived Macrophages for the Study of NO-Redox Biology
Published on: May 31, 2022
1Department of Pediatrics, Xiangya Hospital, Central South University, Changsha 410008, China.
Abstract:
We early show that glutamate (Glu) mediate hyperoxia-induced newborn rat lung injury through N-methyl-D-aspartate receptor (NMDAR). In this study, we search for evidence of NMDAR expression on newborn rat alveolar macrophages (AMs) and the difference between newborn and adult rat AMs, and the possible effect on nitric oxide (NO) production of AMs by exogenous NMDA. The protein of NMDAR was showed by immunocytochemistry, and the mRNA was examined by RT-PCR and real-time PCR. The results show that: (i) both newborn and adult rat AMs express NMDAR1 and the four NMDAR2 subtypes and newborn rat AMs are higher expression. (ii) NMDA administration increase NO production, inducible nitric oxide synthase (iNOS) activity and iNOS mRNA expression of AMs. (iii) NMDAR activation elevates NO secretion of AMs, which suggests that AM may be one of the key cellular origin of the elevated NO secretion in hyperoxia-induced lung injury.
Insights
Glutamate (Glu) and N-methyl-D-aspartate receptors (NMDAR) contribute to newborn rat lung injury. Newborn rat alveolar macrophages (AMs) express higher NMDAR levels, and NMDA increases nitric oxide (NO) production in AMs.
Area of Science:
- Neonatal physiology
- Pulmonary medicine
- Immunology
Background:
- Glutamate (Glu) mediates hyperoxia-induced lung injury in newborn rats via N-methyl-D-aspartate receptors (NMDAR).
- Alveolar macrophages (AMs) play a role in lung injury, but their NMDAR expression and role in nitric oxide (NO) production are not fully understood.
Purpose of the Study:
- To investigate NMDAR expression in newborn and adult rat AMs.
- To determine the effect of exogenous NMDA on NO production in AMs.
- To elucidate the role of NMDAR activation in AM-derived NO secretion.
Main Methods:
- Immunocytochemistry to detect NMDAR protein.
- RT-PCR and real-time PCR to examine NMDAR mRNA expression.
- Assessment of NO production, inducible nitric oxide synthase (iNOS) activity, and iNOS mRNA expression following NMDA administration.
Main Results:
- Both newborn and adult rat AMs express NMDAR1 and NMDAR2 subtypes, with higher expression in newborns.
- NMDA administration significantly increased NO production, iNOS activity, and iNOS mRNA expression in AMs.
- NMDAR activation led to elevated NO secretion from AMs.
Conclusions:
- Newborn rat AMs express higher levels of NMDAR compared to adult AMs.
- NMDAR activation in AMs stimulates NO production, suggesting AMs are a key cellular source of elevated NO in hyperoxia-induced lung injury.
