Sun Y, Deibler G E, Sokoloff L, Smith C B
Laboratory of Cerebral Metabolism, National Institute of Mental Health, Department of Health and Human Services, Bethesda, Maryland 20892.
J Neurochem. 1992 Sep;59(3):863-73. doi: 10.1111/j.1471-4159.1992.tb08324.x.
The quantitative autoradiographic L-[1-14C]leucine method for the determination of regional rates of cerebral protein synthesis in vivo takes into account recycling of unlabeled leucine derived from protein degradation into the precursor pool for protein synthesis. We have evaluated the degree of recycling by measuring the ratio of the apparent steady-state leucine specific activity in the precursor amino acid pool (tRNA-bound leucine) to that in the arterial plasma. In the whole brain of the conscious rat this ratio (lambda WB) equals 0.58. The equivalent ratio for leucine in the acid-soluble pool in whole brain (psi WB) is 0.49. A first-degree polynomial equation for lambda WB as a function of psi WB was fitted from paired determinations. To determine the degree of recycling in local regions of the brain, we have measured in individual brain regions (i) psi i and calculated lambda i assuming that the fitted equation also applies to these localized regions. Our results indicate that the degree of recycling into the precursor pool does vary regionally; lambda i in the individual regions varies from 0.62 in the hypoglossal nucleus to 0.50 in the globus pallidus. Local rates of protein synthesis were then determined by the autoradiographic technique with regional corrections for recycling of unlabeled leucine. Rates of leucine incorporation into protein averaged 6.1 nmol/g of tissue/min in the brain as a whole, with the rates in gray matter about twice those in white matter.
用于测定体内脑区蛋白质合成区域速率的定量放射自显影L-[1-¹⁴C]亮氨酸法考虑了蛋白质降解产生的未标记亮氨酸再循环进入蛋白质合成前体池的情况。我们通过测量前体氨基酸池(与tRNA结合的亮氨酸)中表观稳态亮氨酸比活性与动脉血浆中该比活性的比值来评估再循环程度。在清醒大鼠的全脑中,该比值(λWB)等于0.58。全脑酸溶性池中亮氨酸的等效比值(ψWB)为0.49。根据配对测定结果拟合了λWB作为ψWB函数的一阶多项式方程。为了确定脑局部区域的再循环程度,我们在各个脑区测量了(i)ψi,并假设拟合方程也适用于这些局部区域来计算λi。我们的结果表明,进入前体池的再循环程度确实存在区域差异;各个区域的λi从舌下神经核的0.62到苍白球的0.50不等。然后通过放射自显影技术并对未标记亮氨酸的再循环进行区域校正来确定局部蛋白质合成速率。全脑中亮氨酸掺入蛋白质的速率平均为6.1 nmol/g组织/分钟,灰质中的速率约为白质中的两倍。