Gardiner C S, Williams J S, Menino A R
Department of Animal Science, Oregon State University, Corvallis 97331-6702.
Biol Reprod. 1990 Nov;43(5):788-94. doi: 10.1095/biolreprod43.5.788.
Changes in sodium/potassium adenosine triphosphatase (Na+/K+ ATPase) and Na+/K+ ATPase mRNA content during preimplantation mouse embryo development were determined. Western blotting, using polyclonal antiserum against guinea pig Na+/K+ ATPase, was used to detect changes in Na+/K+ ATPase alpha- and beta-subunit content during mouse embryo development. Total RNA from mouse embryos was analyzed using Northern and slot blots hybridized with random-primer-labeled cDNA for Na+/K+ ATPase alpha-subunit from sheep kidney. Northern blots exhibited a single mRNA band (3.65 kb) in sheep and mouse kidneys and mouse embryos. Although Na+/K+ ATPase alpha-subunit mRNA content of mouse embryos increased 45-fold between Day 1 and Day 4 of development, Na+/K+ ATPase alpha-subunit content remained constant, and beta-subunit content increased 9-fold. The Na+/K+ ATPase alpha-subunit and alpha-subunit mRNA content did not increase in a similar manner. The results suggest that, in mouse embryos, blastocoel formation is not triggered by an increase in Na+/K+ ATPase alpha-subunit content. Changes in beta-subunit content may be important in regulating Na+/K+ ATPase activity and blastocoel formation.
测定了小鼠植入前胚胎发育过程中钠钾腺苷三磷酸酶（Na+/K+ ATP酶）及Na+/K+ ATP酶mRNA含量的变化。利用针对豚鼠Na+/K+ ATP酶的多克隆抗血清进行蛋白质免疫印迹分析，以检测小鼠胚胎发育过程中Na+/K+ ATP酶α和β亚基含量的变化。使用与绵羊肾脏Na+/K+ ATP酶α亚基随机引物标记cDNA杂交的Northern印迹法和狭缝印迹法分析小鼠胚胎的总RNA。Northern印迹显示在绵羊和小鼠肾脏以及小鼠胚胎中均有一条单一的mRNA条带（3.65 kb）。尽管小鼠胚胎的Na+/K+ ATP酶α亚基mRNA含量在发育第1天到第4天之间增加了45倍，但Na+/K+ ATP酶α亚基含量保持恒定，而β亚基含量增加了9倍。Na+/K+ ATP酶α亚基和α亚基mRNA含量的增加方式并不相似。结果表明，在小鼠胚胎中，囊胚腔的形成并非由Na+/K+ ATP酶α亚基含量的增加所触发。β亚基含量的变化可能对调节Na+/K+ ATP酶活性和囊胚腔形成具有重要意义。
