Expression and distribution of N-methyl-D-aspartate receptor 2A/B in anterior thalamic nuclei and hippocampus of rats

Kai Fan; Xiaokai Ma; Bin Wang

摘要：BACKGROUND: Several immunohistochemical and in situ hybridization studies have demonstrated the localization of various N-methyl-D-aspartate receptors (NMDAR) subunits in certain brain regions of mammals. These studies indicate that NMDAR are widespread in mammal brains, especially in cerebral cortex and hippocam pus.OBJECTIVE: To validate whether or not the NMDAR 2A/B localizes in rat anterior thalamic nuclei (ATN) and hippocampus, and observe its expression and distribution characteristics. DESIGN: Controlled study.SETTING: Department of Anatomy, College of Basic Medicine, Dalian Medical University. MATERIALS: Ten adult Sprague-Dawley rats of either gender, weighing 180-250 g, of clean grade, were provided by the Experimental Animal Center, Dalian Medical University. Rabbit polyclonal antibody to NMDAR 2A/B was the product of Chemicon Company.METHODS: This experiment was carried out in the Department of Anatomy, College of Basic Medicine, Dalian Medical University from September 2005 to May 2006. The anesthetized SD rats were transcardially perfused with 100 mL phosphate buffer solution, then perfused with paraformaldehyde through ascending aorta. After 30 minutes, their brains were harvested and post-fixed in PFA at 4 ℃ overnight. Hippocampal and anterior thalamic nuclear tissues were sliced in the coronal plane at 40 μm. The sections were stained by immunohistochemical ABC method, visualized by DAB, dehydrated routinely, performed transparence and coverslipped. Expression and distribution of NMDAR 2A/B in ATN and hippocampus of rats were observed under an optical microscope.MAIN OUTCOME MEASURES: Distribution and expression of NMDAR 2A/B in ATN and hippocampus of rats, RESULTS: NMDAR 2A/B positive neurons localized in ATN, pyramidal layer of hippocampus and granular layer of dentate gyrus in a compact and orderly pattern, and immunostained intensely. NMDAR 2A/B receptor positive neurons scattered with lower intensity of immunolabeling in other regions of hippocampus and dentate gyrus.CONCLUSION: The experimental results suggested that a large number of NMDAR 2A/B localized in ATN, pyramidal layer of hippocampus and granular layer of dentate gyrus in rats.