Liu H C, He Z, Rosenwaks Z
Center for Reproductive Medicine and Infertility, Weill Medical College of Cornell University, New York, New York 10021, USA.
Fertil Steril. 2001 May;75(5):947-55. doi: 10.1016/s0015-0282(01)01706-x.
Using oligonucleotide microarray (DNA chip)-based hybridization analysis to gain a comprehensive view of gene expression and regulation involved in folliculogenesis.
Prospective randomized study.
Academic institution.
ANIMAL(S): B6D2F1 female mice.
INTERVENTION(S): Superovulation.
MAIN OUTCOME MEASURE(S): Preantral follicles isolated from day 14 B6D2F-1 mice were stimulated in vitro to form Graafian follicles. Total RNA extracted from the mouse preantral and Graafian follicles were reverse transcribed, labeled with digoxigenin-11-dUTP, and then hybridized with Clontech Atlas mouse cDNA expression arrays for comparison.
RESULT(S): Of 588 known studied genes, 39 and 61 were detected in preantral follicles and in Graafian follicles, respectively, and 17 were highly expressed consistently in both preantral and Graafian follicles. Performing clustering analysis, we found that 15 detected genes were down-regulated and 46 were up-regulated as the follicles advanced to mature stages.
CONCLUSION(S): We have successfully developed a sensitive DNA chip technology that is able to simultaneously and quantitatively study gene expression profiles in a small number of follicles (1.5-15 follicles). Several folliculogenesis-related genes have been identified. Some of these genes were expressed, indicating that they may be essential for follicle growth and maturation, whereas others were up-regulated only during late follicular development, indicating stage-specific roles.
运用基于寡核苷酸微阵列（DNA芯片）的杂交分析，全面了解卵泡发生过程中涉及的基因表达与调控。
前瞻性随机研究。
学术机构。
B6D2F1雌性小鼠。
超排卵。
从第14天的B6D2F-1小鼠分离出的窦前卵泡在体外进行刺激以形成格拉夫卵泡。从小鼠窦前卵泡和格拉夫卵泡中提取的总RNA进行逆转录，用地高辛-11-dUTP标记，然后与Clontech Atlas小鼠cDNA表达阵列杂交以进行比较。
在588个已知研究基因中，分别在窦前卵泡和格拉夫卵泡中检测到39个和61个，17个在窦前卵泡和格拉夫卵泡中均持续高表达。进行聚类分析时，我们发现随着卵泡发育至成熟阶段，15个检测到的基因下调，46个上调。
我们成功开发了一种灵敏的DNA芯片技术，能够同时定量研究少量卵泡（1.5 - 15个卵泡）中的基因表达谱。已鉴定出几个与卵泡发生相关的基因。其中一些基因有表达，表明它们可能对卵泡生长和成熟至关重要，而其他一些基因仅在卵泡发育后期上调，表明具有阶段特异性作用。
