Frerichs K U, Smith C B, Brenner M, DeGracia D J, Krause G S, Marrone L, Dever T E, Hallenbeck J M
Stroke Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 1998 Nov 24;95(24):14511-6. doi: 10.1073/pnas.95.24.14511.
Protein synthesis (PS) has been considered essential to sustain mammalian life, yet was found to be virtually arrested for weeks in brain and other organs of the hibernating ground squirrel, Spermophilus tridecemlineatus. PS, in vivo, was below the limit of autoradiographic detection in brain sections and, in brain extracts, was determined to be 0.04% of the average rate from active squirrels. Further, it was reduced 3-fold in cell-free extracts from hibernating brain at 37 degreesC, eliminating hypothermia as the only cause for protein synthesis inhibition (active, 0.47 +/- 0.08 pmol/mg protein per min; hibernator, 0.16 +/- 0.05 pmol/mg protein per min, P < 0.001). PS suppression involved blocks of initiation and elongation, and its onset coincided with the early transition phase into hibernation. An increased monosome peak with moderate ribosomal disaggregation in polysome profiles and the greatly increased phosphorylation of eIF2alpha are both consistent with an initiation block in hibernators. The elongation block was demonstrated by a 3-fold increase in ribosomal mean transit times in cell-free extracts from hibernators (active, 2.4 +/- 0.7 min; hibernator, 7.1 +/- 1.4 min, P < 0.001). No abnormalities of ribosomal function or mRNA levels were detected. These findings implicate suppression of PS as a component of the regulated shutdown of cellular function that permits hibernating ground squirrels to tolerate "trickle" blood flow and reduced substrate and oxygen availability. Further study of the factors that control these phenomena may lead to identification of the molecular mechanisms that regulate this state.
蛋白质合成(PS)一直被认为是维持哺乳动物生命所必需的,但研究发现,在冬眠的十三条纹地松鼠(Spermophilus tridecemlineatus)的大脑和其他器官中,蛋白质合成实际上会停止数周。在体内,大脑切片中的蛋白质合成低于放射自显影检测的极限,并且在脑提取物中,其水平被确定为活跃松鼠平均速率的0.04%。此外,在37摄氏度下,冬眠大脑的无细胞提取物中的蛋白质合成速率降低了3倍,这排除了体温过低是蛋白质合成抑制的唯一原因(活跃状态下,每分钟每毫克蛋白质为0.47±0.08皮摩尔;冬眠状态下,每分钟每毫克蛋白质为0.16±0.05皮摩尔,P<0.001)。蛋白质合成的抑制涉及起始和延伸阶段的阻断,其开始与进入冬眠的早期过渡阶段相吻合。多核糖体图谱中单体峰增加且核糖体有适度解聚,以及真核起始因子2α(eIF2α)磷酸化大幅增加,这两者都与冬眠动物的起始阻断一致。通过冬眠动物无细胞提取物中核糖体平均转运时间增加3倍证明了延伸阻断(活跃状态下,2.4±0.7分钟;冬眠状态下,7.1±1.4分钟,P<0.001)。未检测到核糖体功能或mRNA水平的异常。这些发现表明,蛋白质合成的抑制是细胞功能调节性关闭的一个组成部分,这使得冬眠的十三条纹地松鼠能够耐受“涓涓细流”般的血流以及底物和氧气供应的减少。对控制这些现象的因素进行进一步研究,可能会导致识别出调节这种状态的分子机制。