A new stable isotope method enables the simultaneous measurement of nucleic acid and protein synthesis in vivo in mice.
A new stable isotope method enables the simultaneous measurement of nucleic acid and protein synthesis in vivo in mice.
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一种新的稳定同位素方法能够同时测量小鼠体内核酸和蛋白质的合成。
DOI：
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
P. Reeds
中科院分区：
文献类型：
--
作者：
J. F. Pérez;P. Reeds
我们开发了一种方法的基础上纳入的13 C2-单位来自[U-13 C]甘氨酸，允许同时定量的组织蛋白质和RNA的合成在体内。给两组26只小鼠喂食含有高纤维（HF饮食）或低纤维量（LF饮食）的饮食。6天后，将[U13 C]甘氨酸添加到饲料中，并在2、4、6、8、12和24小时后杀死每组四只小鼠。提取肝和肠粘膜游离和RNA结合的嘌呤核苷，并酶降解为尿囊素。尿囊素降解为乙醛酸，然后还原胺化为甘氨酸，甘氨酸含有通过从头合成并入的两个13 C原子。HF饮食的摄入与肝脏（HF饮食，29%/d; LF饮食，21%/d）和粘膜（HF饮食，27%/d; LF饮食，17%/d）中显著（P < 0.05）更高的总RNA合成速率相关。各组织中RNA合成的平均速率显著（P < 0.01）低于各自的蛋白质合成速率（肝脏，67%/d;粘膜，74%/d）。游离嘌呤核苷酸库的同位素富集度呈指数增长，但稳态值显著低于正常值（P < 0. 05）。001)低于RNA结合的嘌呤。结果表明，游离核苷酸池由两个动力学不同的隔间，其中一个是小的，并具有快速的周转率。我们建议，这是作为RNA前体库。另一个是大的，有一个低的周转率，我们相信，是涉及细胞能量的腺苷三磷酸池。
We developed a method based on the incorporation of 13C2-units derived from [U-13C]glycine that allows the simultaneous quantification of tissue protein and RNA synthesis in vivo. Two groups of 26 mice were fed diets containing a high (HF diet) or a low quantity of fiber (LF diet). After 6 d, [U13C]glycine was added to the diet and groups of four mice were killed after 2, 4, 6, 8, 12 and 24 h. Hepatic and intestinal mucosal free and RNA-bound purine nucleosides were extracted and enzymically degraded to allantoin. Allantoin was degraded to glyoxylate, which was then reductively aminated to glycine, which contains the two 13C-atoms incorporated via de novo synthesis. Ingestion of the HF diet was associated with significantly (P < 0.05) higher rates of total RNA synthesis in both the liver ( HF diet, 29%/d; LF diet, 21%/d) and mucosa (HF diet, 27%/d; LF diet, 17 %/d). The mean rates of RNA synthesis in each tissue were significantly (P < 0.01) lower than the respective rates of protein synthesis (liver, 67%/d; mucosa, 74%/d). The isotopic enrichment of the free purine nucleotide pool increased rapidly and exponentially, but the steady-state value was substantially (P < 0. 001) lower than that of the RNA-bound purines. The results suggest that the free nucleotide pool consists of two kinetically distinct compartments, one of which is small and has a rapid rate of turnover. This, we propose, acts as the RNA precursor pool. The other is large, has a low rate of turnover and, we believe, is the pool of adenosine triphosphate involved in cellular energetics.
幼猪血浆蛋白合成率的测量：替代示踪剂方法的研究。
DOI：
10.1152/ajpregu.1994.267.1.r221
发表时间：
1994
期刊：
The American journal of physiology
影响因子：
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作者：
Jahoor,F;Burrin,DG;Reeds,PJ;Frazer,M
通讯作者：
Frazer,M
DOI：
10.1016/s0021-9258(18)47824-6
发表时间：
1987-10
期刊：
The Journal of biological chemistry
影响因子：
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作者：
B. Lardeux;S. Heydrick;G. E. Mortimore
通讯作者：
B. Lardeux;S. Heydrick;G. E. Mortimore
饮食量对大鼠小肠形态和细胞更新的影响。
影响因子：
29.4
作者：
Ecknauer,R;Sircar,B;Johnson,LR
通讯作者：
Johnson,LR