Ace C I, Okulicz W C
Departments of OB/GYN and Physiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
Biol Reprod. 1999 Apr;60(4):1029-35. doi: 10.1095/biolreprod60.4.1029.
We used differential-display reverse transcription-polymerase chain reaction (DDRT-PCR) to identify different patterns of progesterone (P4)-dependent gene regulation in rhesus monkey endometria. Complementary DNA populations representing the proliferative phase (estrogen dominant, EcDNA) and an inadequate secretory phase (low level of P4, IcDNA) were compared with a cDNA population representing an adequate secretory phase (normal level of P4, PcDNA). We were able to distinguish four different levels of mRNA regulation: 1) up-regulation by P4 during an adequate secretory phase, 2) autologous down-regulation (IcDNA versus PcDNA), 3) lower abundance in IcDNA compared to PcDNA, and 4) P4-dependent inhibition of EcDNA gene expression. We isolated and sequenced 16 fragments representing these different levels of P4 regulation. The sequence of three fragments that were autologously down-regulated (I1, I2, I4) matched previously entered GenBank mRNAs: I1 encodes serine/threonine protein phosphatase A; I2 encodes oxobutanoate dehydrogenase E1b-beta; and I4 encodes line-1 reverse transcriptase homologue. Six other fragments exhibited homology to uncharacterized expressed sequence tags, sequence site tags, and cosmid clones. The remaining seven fragments exhibited no significant homology to GenBank entries at this time. The various patterns of P4-dependent gene regulation identified in the present study are likely to play roles in the temporal orchestration of events that lead to proper maturation of the endometrium.
我们使用差异显示逆转录-聚合酶链反应(DDRT-PCR)来鉴定恒河猴子宫内膜中孕酮(P4)依赖性基因调控的不同模式。将代表增殖期(雌激素占主导,EcDNA)和分泌期不足(P4水平低,IcDNA)的互补DNA群体与代表分泌期充足(P4水平正常,PcDNA)的cDNA群体进行比较。我们能够区分四种不同水平的mRNA调控:1)在分泌期充足时P4上调;2)自体下调(IcDNA与PcDNA相比);3)与PcDNA相比,IcDNA中丰度较低;4)P4依赖性抑制EcDNA基因表达。我们分离并测序了代表这些不同P4调控水平的16个片段。三个自体下调片段(I1、I2、I4)的序列与先前录入GenBank的mRNA匹配:I1编码丝氨酸/苏氨酸蛋白磷酸酶A;I2编码氧代丁酸脱氢酶E1b-β;I4编码1型逆转录酶同源物。其他六个片段与未鉴定的表达序列标签、序列位点标签和粘粒克隆具有同源性。其余七个片段目前与GenBank条目无明显同源性。本研究中鉴定出的P4依赖性基因调控的各种模式可能在导致子宫内膜正常成熟的事件的时间编排中发挥作用。