Jiang P, Arinze I J
Department of Biochemistry, Meharry Medical College, Nashville, TN 37208.
Mol Cell Endocrinol. 1994 Feb;99(1):95-102.
The mRNA levels for Gs alpha and G beta in liver from various age groups of rabbits were assessed by Northern and dot-blot hybridization assays using cDNA and oligonucleotide probes. The mRNA levels for both Gs alpha and G beta exhibited a transient 30-35% decrease at 3-6 h after birth, followed by a 3- to 3.5-fold increase which peaked at 2 days after birth, then gradually declined to adult levels at 4-6 weeks. The changes in mRNA levels paralleled the changes in protein levels [previously measured by immunoblotting (Kawai and Arinze, 1991)] in the first 2 days after birth, but not in the later periods of development. Beyond day 3, the increase in protein levels persists, reaching maximal levels at 4 weeks and equalling adult levels while their mRNA levels decrease to about 50% of the levels at day 2. In vivo administration of dexamethasone to neonatal rabbits increased hepatic mRNA levels for both Gs alpha and G beta by about 50%; the effect was moderate compared to the 2- to 3-fold increase in corresponding protein levels. The transcription rate for the Gs alpha gene increased by only 32% at day 2 after birth compared to term. A 68% increase in transcription rate for this gene was observed after the dexamethasone treatment. Taken together, these data indicate that the developmental and glucocorticoid-regulated expression of Gs alpha- and G beta-subunits in neonatal liver is modulated at the transcriptional level.
利用cDNA和寡核苷酸探针,通过Northern印迹法和斑点杂交法评估了不同年龄组兔肝脏中Gsα和Gβ的mRNA水平。Gsα和Gβ的mRNA水平在出生后3 - 6小时均出现短暂下降,降幅为30 - 35%,随后在出生后2天增加3 - 3.5倍并达到峰值,然后在4 - 6周逐渐降至成年水平。在出生后的前两天,mRNA水平的变化与蛋白质水平的变化(先前通过免疫印迹法测定,Kawai和Arinze,1991)平行,但在发育后期则不然。出生3天后,蛋白质水平持续增加,在4周时达到最高水平并与成年水平相当,而它们的mRNA水平降至出生后第2天水平的约50%。对新生兔进行地塞米松的体内给药使肝脏中Gsα和Gβ的mRNA水平均增加约50%;与相应蛋白质水平2 - 3倍的增加相比,该效应较为适度。与足月时相比,出生后第2天Gsα基因的转录率仅增加32%。地塞米松治疗后,该基因的转录率增加了68%。综上所述,这些数据表明新生兔肝脏中Gsα和Gβ亚基的发育和糖皮质激素调节表达在转录水平上受到调控。