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通过离体1H(13C)核磁共振波谱检测发育中的大脑皮质中的葡萄糖代谢。

Glucose metabolism in the developing cerebral cortex as detected by 1H(13C) nuclear magnetic resonance spectroscopy ex vivo.

作者信息

Pirttilä T R, Auriola S O, Kauppinen R A

机构信息

Department of Biochemistry and Biotechnology, A. I. Virtanen Institute, Kuopio, Finland.

出版信息

J Neurochem. 1995 Jan;64(1):417-23. doi: 10.1046/j.1471-4159.1995.64010417.x.

DOI:10.1046/j.1471-4159.1995.64010417.x
PMID:7798941
Abstract

Metabolism of [1-13C]glucose was monitored in superfused cerebral cortex slice preparations from 1-, 2-, and 5-week-old rats using 1H-observed/13C-edited (1H(13C)) NMR spectroscopy. The rate of label incorporation into glutamate C-4 did not differ among the three age groups: 0.52-0.67% of total 1H NMR-detected glutamate/min. This was rather unexpected, as oxygen uptake proceeded at 1.1 +/- 0.1, 1.9 +/- 0.1, and 2.0 +/- 0.1 mumol/min/g wet weight in brain slices prepared from 1-, 2-, and 5-week-old animals, respectively. Steady-state glutamate C-4 fractional enrichments in the slice preparations were approximately 23% in all age groups. In the acid extracts of slices glutamate C-4 enrichments were smaller, however, in 1- and 2-week-old (17.8 +/- 1.7 and 16.8 +/- 0.8%, respectively) than in 5-week-old rats (22.7 +/- 0.7%) after 75 min of incubation with 5 mM [1-13C]glucose. We add a new assignment to the 1H(13C) NMR spectroscopy, as acetate C-2 was detected in slice preparations from 5-week-old animals. In the acid extracts of slice preparations acetate C-2 was labeled by approximately 30% in 5-week-old rats but by 15% in both 1- and 2-week-old animals, showing that the turnover rate was increased in 5-week-old animals. In the extracts 3-4% of the C-6 of N-acetyl-aspartate (NAA; CH3 of the acetyl group) contained label as determined by both NMR and mass spectrometry, which indicated that there was no significant labeling to other carbons in NAA.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

采用氢观测/碳编辑(1H(13C))核磁共振波谱法,监测了1周龄、2周龄和5周龄大鼠脑片制备物中[1-13C]葡萄糖的代谢情况。三个年龄组中,标记物掺入谷氨酸C-4的速率没有差异:占1H核磁共振检测到的总谷氨酸的0.52-0.67%/分钟。这相当出乎意料,因为在分别由1周龄、2周龄和5周龄动物制备的脑片中,氧摄取速率分别为1.1±0.1、1.9±0.1和2.0±0.1微摩尔/分钟/克湿重。在所有年龄组的脑片制备物中,稳态谷氨酸C-4的丰度约为23%。然而,在脑片的酸提取物中,与5 mM [1-13C]葡萄糖孵育75分钟后,1周龄和2周龄大鼠(分别为17.8±1.7%和16.8±0.8%)的谷氨酸C-4丰度低于5周龄大鼠(22.7±0.7%)。我们为1H(13C)核磁共振波谱法增加了一项新的归属,因为在5周龄动物的脑片制备物中检测到了乙酸盐C-2。在脑片制备物的酸提取物中,5周龄大鼠的乙酸盐C-2标记率约为30%,而1周龄和2周龄动物均为15%,表明5周龄动物的周转率有所增加。在提取物中,通过核磁共振和质谱测定,N-乙酰天门冬氨酸(NAA;乙酰基的CH3)的C-6中有3-4%含有标记物,这表明NAA的其他碳没有明显标记。(摘要截断于250字)

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