Sun Y, Deibler G E, Smith C B
Laboratory of Cerebral Metabolism, National Institute of Mental Health, Bethesda, MD 20892.
J Cereb Blood Flow Metab. 1993 Nov;13(6):1006-12. doi: 10.1038/jcbfm.1993.126.
The quantitative autoradiographic L-[1-14C] leucine method for the determination of regional rates of cerebral protein synthesis (lCPSleu) requires knowledge of the degree of recycling of leucine derived from protein degradation into the precursor pool for protein synthesis, which can be evaluated by measuring lambda i, the steady-state ratio of the leucine-specific activity in the precursor amino acid pool (tRNA-bound leucine) to that of the arterial plasma. To define the changes in lCPSleu during regeneration of the hypoglossal nerve, we examined the effects of axotomy on the value of lambda i. Because the concentration of tRNA-bound leucine in the hypoglossal nucleus is too low to measure, we measured the equivalent ratio for the total acid-soluble pool (psi i) and applied the linear relationship between lambda and psi found in the whole brain to calculate a value of lambda i in the ipsilateral and contralateral hypoglossal nuclei of 22 adult female rats 2, 18, 35, and 60 days after unilateral hypoglossal axotomy. Statistically significant but quantitatively inconsequential effects of axotomy on values of psi i and lambda i were found. Therefore, the mean value for lambda i (0.64) of the left and right hypoglossal nuclei in all 22 axotomized rats was used to calculate lCPSleu. In a separate group of 15 unilaterally axotomized rats, lCPSleu was determined by the autoradiographic technique; lCPSleu was increased on the axotomized side by 23% on day 2, 30% on day 18, and 13% on day 35. By postaxotomy day 60, lCPSleu had returned to normal.
用于测定脑蛋白质合成区域速率(lCPSleu)的定量放射自显影L-[1-14C]亮氨酸法,需要了解源自蛋白质降解的亮氨酸再循环进入蛋白质合成前体池的程度,这可以通过测量λi来评估,λi是前体氨基酸池(tRNA结合的亮氨酸)中亮氨酸比活性与动脉血浆中亮氨酸比活性的稳态比值。为了确定舌下神经再生过程中lCPSleu的变化,我们研究了轴突切断对λi值的影响。由于舌下神经核中tRNA结合的亮氨酸浓度过低无法测量,我们测量了总酸溶性池的等效比值(ψi),并应用全脑中发现的λ与ψ之间的线性关系,计算了22只成年雌性大鼠单侧舌下神经切断后2、18、35和60天同侧和对侧舌下神经核中的λi值。发现轴突切断对ψi和λi值有统计学上显著但数量上无关紧要的影响。因此,在所有22只切断轴突的大鼠中,左右舌下神经核的λi平均值(0.64)用于计算lCPSleu。在另一组15只单侧切断轴突的大鼠中,通过放射自显影技术测定lCPSleu;在切断轴突的一侧,lCPSleu在第2天增加了23%,在第18天增加了30%,在第35天增加了13%。到轴突切断后第60天,lCPSleu已恢复正常。