Franklin S O, Elliott K, Zhu Y S, Wahlestedt C, Inturrisi C E
Department of Pharmacology, Cornell University Medical College, New York, NY 10021.
Brain Res Mol Brain Res. 1993 Jul;19(1-2):93-100. doi: 10.1016/0169-328x(93)90153-g.
A rapid and sensitive solution hybridization assay was used to quantitate N-methyl-D-aspartate (NMDA) receptor mRNA levels in the central nervous system (CNS) of rat, mouse and human. A riboprobe labelled with 32P was prepared from a plasmid containing a 1413 base sequence from the cDNA for the functional rat NMDA receptor subunit, NMDAR1. Using a full length sense transcript as the calibration standard, the assay reliably measures 8 pg of NMDAR1 mRNA. When expressed as pg of NMDAR1 mRNA/micrograms total cellular RNA, the highest levels in the adult rat CNS are in the olfactory bulb (20.9 pg/micrograms RNA) and the lowest levels are in the spinal cord (5.2 pg/micrograms RNA). Intermediate levels were found in frontal cortex, hippocampus, cerebellum and whole brain. In the mouse CNS the highest levels of NMDAR1 mRNA were found in the olfactory bulb (12.9 pg equivalents/micrograms RNA), followed closely by hippocampus, frontal cortex and cerebellum. Mouse spinal cord (4.4 pg equivalents/micrograms RNA) had the lowest levels of NMDAR1 mRNA. The NMDAR1 riboprobe hybridizes with the same size transcripts in Poly(A)+ RNA from rat, mouse and human brain. In the developing rat, NMDAR1 mRNA levels in frontal cortex and hippocampus increased nearly 3 fold from postnatal day 3 to day 15 and approximately doubled from day 15 to day 67 (adult). Therefore, from postnatal day 3 to adult (day 67) frontal cortex and hippocampus levels of NMDAR1 mRNA increased nearly 6 fold.(ABSTRACT TRUNCATED AT 250 WORDS)
采用一种快速灵敏的溶液杂交分析法，对大鼠、小鼠和人类中枢神经系统（CNS）中N-甲基-D-天冬氨酸（NMDA）受体mRNA水平进行定量。从含有功能性大鼠NMDA受体亚基NMDAR1的cDNA的1413个碱基序列的质粒制备用32P标记的核糖探针。以全长正义转录本作为校准标准，该分析法可可靠地检测8 pg的NMDAR1 mRNA。当以pg NMDAR1 mRNA/微克总细胞RNA表示时，成年大鼠CNS中最高水平在嗅球（20.9 pg/微克RNA），最低水平在脊髓（5.2 pg/微克RNA）。在额叶皮质、海马体、小脑和全脑中发现的水平处于中间。在小鼠CNS中，NMDAR1 mRNA的最高水平在嗅球（12.9 pg当量/微克RNA），其次是海马体、额叶皮质和小脑。小鼠脊髓（4.4 pg当量/微克RNA）中NMDAR1 mRNA水平最低。NMDAR1核糖探针与大鼠、小鼠和人类大脑的Poly(A)+ RNA中相同大小的转录本杂交。在发育中的大鼠中，额叶皮质和海马体中NMDAR1 mRNA水平从出生后第3天到第15天增加近3倍，从第15天到第67天（成年）大约翻倍。因此，从出生后第3天到成年（第67天），额叶皮质和海马体中NMDAR1 mRNA水平增加近6倍。（摘要截短于250字）
