Inserm, U625, Université Rennes 1, IFR140, Rennes, F-35042, France.
Reprod Biol Endocrinol. 2009 Nov 24;7:133. doi: 10.1186/1477-7827-7-133.
The LIM domain protein Fhl5 was previously found to interact with CREM, a DNA binding transcriptional regulator necessary for spermiogenesis in mammals. Co-transfection experiments using heterologous promoter constructs indicated a role for Fhl5 in transcriptional up-regulation of CREM-dependent testicular genes. Male mice lacking Fhl5 were reported to be fertile but displayed partially abnormal sperm maturation and morphology.
To identify Fhl5 testicular target genes we carried out two whole-genome expression profiling experiments using high-density oligonucleotide microarrays and total testis samples from Fhl5 wild-type versus homozygous mutant mice first in different and then in isogenic strain backgrounds.
Weak signal differences were detected in non-isogenic samples but no statistically significant expression changes were observed when isogenic Fhl5 mutant and wild-type samples were compared.
The outcome of these experiments suggests that testicular expression profiling is extremely sensitive to the genetic background and that Fhl5 is not essential for testicular gene expression to a level detected by microarray-based measurements. This might be due to redundant function of the related and similarly expressed protein Fhl4.
LIM 结构域蛋白 Fhl5 先前被发现与 CREM 相互作用,CREM 是一种 DNA 结合转录调节因子,对于哺乳动物的精子发生是必需的。使用异源启动子构建体的共转染实验表明,Fhl5 在 CREM 依赖性睾丸基因的转录上调中发挥作用。报道称缺乏 Fhl5 的雄性小鼠具有生育能力,但表现出部分异常的精子成熟和形态。
为了鉴定 Fhl5 的睾丸靶基因,我们使用高密度寡核苷酸微阵列进行了两项全基因组表达谱实验,并使用 Fhl5 野生型与纯合突变型小鼠的睾丸样本进行了实验,首先是在不同的然后是在同基因系背景下进行的。
在非同基因系样本中检测到弱信号差异,但当比较同基因 Fhl5 突变型和野生型样本时,没有观察到统计学上显著的表达变化。
这些实验的结果表明,睾丸表达谱分析对遗传背景非常敏感,并且 Fhl5 对于通过基于微阵列的测量检测到的睾丸基因表达不是必需的。这可能是由于相关且表达相似的蛋白 Fhl4 的冗余功能所致。