Bartels H, Herbort H, Jungermann K
Institut für Biochemie, Georg-August-Universität, Göttingen, Federal Republic of Germany.
Histochemistry. 1990;94(6):637-44. doi: 10.1007/BF00271991.
The zonal distribution of phosphoenolpyruvate carboxykinase (PCK) and tyrosine aminotransferase (TAT) mRNA in liver was studied by in situ hybridization with radiolabelled cRNA probes and the abundance of PCK and TAT mRNA was quantified by Northern blot analysis of total RNA with biotinylated cRNA probes. Livers were taken from rats during a normal 12 h day/night rhythm, when they had access to food only during the dark period from 7 pm to 7 am, or during refeeding, when they had access to food after having been starved for 60 h. 1. Daily feeding rhythm: High levels of PCK mRNA were distributed mainly in the periportal and intermediate zone during the fasting period at noon and 6 pm. Feeding caused a rapid decrease in PCK mRNA level and a restriction of PCK mRNA localization to the periportal area within the first 2 h. No further alterations were observed during the following hours of the feeding period. TAT mRNA was distributed also in the periportal and intermediate zone during the fasting period. Feeding first reduced the mRNA level without changing the distribution pattern. Then towards the end of the feeding period TAT mRNA increased again to half-maximal levels and became restricted mainly to the periportal area. 2. Starvation-refeeding cycle: High amounts of PCK mRNA as well as of TAT mRNA were localized predominantly in the periportal and intermediate zone after 60 h of starvation. PCK and TAT mRNA both decreased markedly during the first 2 h of refeeding and then remained almost constant.(ABSTRACT TRUNCATED AT 250 WORDS)
通过用放射性标记的cRNA探针进行原位杂交研究了肝脏中磷酸烯醇丙酮酸羧激酶(PCK)和酪氨酸转氨酶(TAT)mRNA的区域分布,并通过用生物素化的cRNA探针进行总RNA的Northern印迹分析来定量PCK和TAT mRNA的丰度。从大鼠身上获取肝脏,这些大鼠处于正常的12小时昼夜节律中,即它们仅在晚上7点到早上7点的黑暗期进食,或者在重新喂食期间,即在饥饿60小时后进食。1. 每日进食节律:中午和下午6点禁食期间,高水平的PCK mRNA主要分布在门静脉周围和中间区域。进食导致PCK mRNA水平迅速下降,并在最初2小时内将PCK mRNA定位限制在门静脉周围区域。在进食期的接下来几个小时内未观察到进一步变化。禁食期间,TAT mRNA也分布在门静脉周围和中间区域。进食首先降低了mRNA水平,而不改变分布模式。然后在进食期结束时,TAT mRNA再次增加到最大水平的一半,并主要局限于门静脉周围区域。2. 饥饿-再喂养周期:饥饿60小时后,大量的PCK mRNA以及TAT mRNA主要定位于门静脉周围和中间区域。再喂养的最初2小时内,PCK和TAT mRNA均显著下降,然后几乎保持不变。(摘要截短于250字)