Department of Reproductive Pharmacology, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
Reprod Biol Endocrinol. 2010 Apr 22;8:37. doi: 10.1186/1477-7827-8-37.
As one of the chlorinated antifertility compounds, alpha-chlorohydrin (ACH) can inhibit glyceraldehyde-3-phosphate dehydrogenase (G3PDH) activity in epididymal sperm and affect sperm energy metabolism, maturation and fertilization, eventually leading to male infertility. Further studies demonstrated that the inhibitory effect of ACH on G3PDH is not only confined to epididymal sperm but also to the epididymis. Moreover, little investigation on gene expression changes in the epididymis after ACH treatment has been conducted. Therefore, gene expression studies may indicate new epididymal targets related to sperm maturation and fertility through the analysis of ACH-treated infertile animals.
Rats were treated with ACH for ten consecutive days, and then each male rat copulated with two female rats in proestrus. Then sperm maturation and other fertility parameters were analyzed. Furthermore, we identified epididymal-specific genes that are associated with fertility between control and ACH groups using an Affymetrix Rat 230 2.0 oligo-microarray. Finally, we performed RT-PCR analysis for several differentially expressed genes to validate the alteration in gene expression observed by oligonucleotide microarray.
Among all the differentially expressed genes, we analyzed and screened the down-regulated genes associated with metabolism processes, which are considered the major targets of ACH action. Simultaneously, the genes that were up-regulated by chlorohydrin were detected. The genes that negatively regulate sperm maturation and fertility include apoptosis and immune-related genes and have not been reported previously. The overall results of PCR analysis for selected genes were consistent with the array data.
In this study, we have described the genome-wide profiles of gene expression in the epididymides of infertile rats induced by ACH, which could become potential epididymal specific targets for male contraception and infertility treatment.
α-氯醇（ACH）作为一种氯化抗生育化合物，可以抑制附睾精子中的甘油醛-3-磷酸脱氢酶（G3PDH）活性，影响精子能量代谢、成熟和受精，最终导致男性不育。进一步的研究表明，ACH 对 G3PDH 的抑制作用不仅局限于附睾精子，还局限于附睾。此外，对 ACH 处理后附睾基因表达变化的研究甚少。因此，通过分析 ACH 处理后的不育动物的基因表达研究，可能会发现与精子成熟和生育力相关的新的附睾靶标。
大鼠连续 10 天用 ACH 处理，然后每只雄性大鼠与两只动情前期的雌性大鼠交配。然后分析精子成熟和其他生育参数。此外，我们使用 Affymetrix Rat 230 2.0 寡聚微阵列，鉴定了与对照组和 ACH 组之间生育力相关的附睾特异性基因。最后，我们对几个差异表达基因进行 RT-PCR 分析，以验证寡核苷酸微阵列观察到的基因表达变化。
在所有差异表达的基因中，我们分析并筛选了与代谢过程相关的下调基因，这些基因被认为是 ACH 作用的主要靶标。同时，还检测到了被氯醇上调的基因。负调控精子成熟和生育力的基因包括凋亡和免疫相关基因，这些基因以前没有报道过。所选基因的 PCR 分析的总体结果与数组数据一致。
在这项研究中，我们描述了 ACH 诱导不育大鼠附睾基因表达的全基因组图谱，这些图谱可能成为男性避孕和不育治疗的潜在附睾特异性靶标。
