Bittel Douglas C, Kibiryeva Nataliya, McNulty Steven G, Driscoll Daniel J, Butler Merlin G, White Robert A
Children's Mercy Hospitals and Clinics and University of Missouri-Kansas City, School of Medicine, Kansas City, Missouri 54108, USA.
Am J Med Genet A. 2007 Mar 1;143A(5):422-9. doi: 10.1002/ajmg.a.31504.
Prader-Willi syndrome (PWS) is caused by loss of paternally expressed genes in the 15q11-q13 region. To further characterize alterations in gene expression in this classical obesity syndrome we used whole genome microarrays to study a PWS mouse model resulting from a paternally derived imprinting center (IC) deletion (PWS IC deletion). These mice die generally within 2-3 days of life (reflective of failure to thrive in infants with PWS) and therefore, the analysis was performed on RNA extracted from the whole brain of PWS IC deletion mice and normal littermates at less than 24 hr after birth. Of more than 45,000 probes examined, 26,471 (59%) were detected for further analysis, and 69 had a significant change in expression of at least 1.5-fold and a false discovery rate (FDR) of 5%. Eight of the genes with differential expression were imprinted and from the PWS critical region (PWSCR). The three genes with the highest expression in the PWS IC mice were pro-opiomelanocortin (Pomc) and two transcripts of unknown function. Pomc knockout mice have been shown to develop obesity. Therefore, elevated Pomc RNA in PWS IC deletion neonatal mice may be an important genetic factor in the survival of these mice as it may affect eating behavior. Interestingly, Mc5r, a melanocortin receptor known to directly respond to Pomc expression changes, was upregulated as well. Mc5r is known to be involved with thermoregulation which is reportedly abnormal in PWS infants. These observations support a role for Pomc and the network of genes involved in regulating energy homeostasis in the early clinical findings of failure to thrive observed in PWS. Other notable patterns include three previously unstudied transcripts that are expressed only from the paternal allele under regulatory control of the IC and include AK013560, BB3144814, and BB182944 (whose genes are located in the mouse PWSCR on chromosome 7B). As expected, all the known paternally expressed genes from the PWSCR had detection signals below the threshold in the PWS IC deletion mice but were clearly detectable in control littermates. Several of the genes in this study were further examined by quantitative reverse transcription-PCR (RT-PCR) to confirm their expression status. Further analysis of gene expression in these mice may lead to novel pathways affected in PWS. These results, along with other recent reports, suggest that the cumulative effect of modest changes in expression of many genes, especially genes involved in energy metabolism, contribute to the failure to thrive of infants with PWS.
普拉德-威利综合征(PWS)是由15号染色体q11-q13区域父源表达基因的缺失所致。为了进一步明确这种典型肥胖综合征中基因表达的改变,我们使用全基因组微阵列研究了一种因父源印记中心(IC)缺失导致的PWS小鼠模型(PWS IC缺失)。这些小鼠通常在出生后2 - 3天内死亡(这反映了PWS婴儿的生长发育不良),因此,分析是在出生后不到24小时从PWS IC缺失小鼠和正常同窝小鼠的全脑中提取的RNA上进行的。在检测的超过45,000个探针中,有26,471个(59%)被检测到用于进一步分析,其中69个的表达有至少1.5倍的显著变化且错误发现率(FDR)为5%。差异表达的基因中有8个是印记基因且来自PWS关键区域(PWSCR)。在PWS IC小鼠中表达最高的三个基因是促阿片黑素皮质素原(Pomc)和两个功能未知的转录本。已证明Pomc基因敲除小鼠会发生肥胖。因此,PWS IC缺失新生小鼠中Pomc RNA升高可能是这些小鼠存活的一个重要遗传因素,因为它可能影响进食行为。有趣的是,黑素皮质素受体5(Mc5r),一种已知直接对Pomc表达变化作出反应的受体,也上调了。已知Mc5r参与体温调节,据报道PWS婴儿存在体温调节异常。这些观察结果支持了Pomc以及参与调节能量稳态的基因网络在PWS中观察到的早期生长发育不良临床发现中的作用。其他值得注意的模式包括三个以前未研究过的转录本,它们仅在IC的调控下从父本等位基因表达,包括AK013560、BB3144814和BB182944(其基因位于小鼠7B染色体上的PWSCR中)。正如预期的那样,来自PWSCR的所有已知父源表达基因在PWS IC缺失小鼠中的检测信号低于阈值,但在对照同窝小鼠中可清晰检测到。本研究中的几个基因通过定量逆转录PCR(RT-PCR)进行了进一步检测以确认其表达状态。对这些小鼠基因表达的进一步分析可能会揭示PWS中受影响的新途径。这些结果,连同最近的其他报道,表明许多基因表达的适度变化的累积效应,特别是参与能量代谢的基因,导致了PWS婴儿的生长发育不良。