Developmental expression and function of DKKL1/Dkkl1 in humans and mice.
Developmental expression and function of DKKL1/Dkkl1 in humans and mice.
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DKKL1/Dkkl1 在人和小鼠中的发育表达和功能
DOI：
10.1186/1477-7827-10-51
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发表时间：
2012-07-21
期刊：
影响因子：
--
通讯作者：
Cai Z
中科院分区：
文献类型：
--
作者：
Yan Q;Wu X;Chen C;Diao R;Lai Y;Huang J;Chen J;Yu Z;Gui Y;Tang A;Cai Z
背景
实验旨在鉴定Dickkopf-Like 1（DKKL 1/Dkkl 1）基因在人类和小鼠中的发育表达和功能。
方法
在多个出生后时间（第4、9、18、35和54天，以及6个月）收集小鼠睾丸cDNA样品，并与Affyssin小鼠全基因组基因芯片杂交。为了进一步研究DKKL 1在人类中的同源基因，我们利用Affytelium人类基因芯片比较了DKKL 1在人类成年睾丸和胎儿睾丸中的表达谱。利用各种细胞和分子生物学技术分析DKKL 1/Dkkl 1的特性。
结果
RT-PCR和Western blot结果证实，Dkkl 1在第4天和第9天的小鼠睾丸中未检测到表达，但在第18、35和54天以及6个月时存在。对Dkkl 1的组织分布的检查表明，虽然Dkkl 1 mRNA在睾丸中大量表达，但在附睾或其他组织中几乎没有观察到Dkkl 1的表达。在体外受精试验中，发现Dkkl 1抗体显著减少受精。人类基因芯片结果显示，DKKL 1在成人睾丸中的杂交信号强度是在胎儿睾丸中的405.56倍。多个人体组织的RT-PCR分析表明，DKKL 1 mRNA仅在睾丸中表达。Western blot分析也表明DKKL 1主要在人睾丸中表达，分子量约为34 kDa。此外，免疫组织化学染色显示，DKKL 1蛋白主要位于精母细胞和圆形精子细胞在人类睾丸。DKKL 1在不育男性患者中的表达水平的检查显示，虽然在仅支持细胞综合征（SCOS）或隐睾症患者的睾丸中没有DKKL 1出现，但在生精停滞患者中观察到DKKL 1的可变表达。
结论
这些结果与以前的研究一起表明，DKKL 1/Dkkl 1可能在睾丸发育和精子发生中起重要作用，并且可能是男性不育的重要因素。
Background
Experiments were designed to identify the developmental expression and function of the Dickkopf-Like1 (DKKL1/Dkkl1) gene in humans and mice.
Methods
Mouse testes cDNA samples were collected at multiple postnatal times (days 4, 9, 18, 35, and 54, as well as at 6 months) and hybridized to Affymetrix mouse whole genome Genechips. To further characterize the homologous gene DKKL1 in human beings, the expression profiles between human adult testis and foetal testis were compared using Affymetrix human Genechips. The characteristics of DKKL1/Dkkl1 were analysed using various cellular and molecular biotechnologies.
Results
The expression of Dkkl1 was not detected in mouse testes on days 4 or 9, but was present on days 18, 35, and 54, as well as at 6 months, which was confirmed by RT-PCR and Western blot results. Examination of the tissue distribution of Dkkl1 demonstrated that while Dkkl1 mRNA was abundantly expressed in testes, little to no expression of Dkkl1 was observed in the epididymis or other tissues. In an in vitro fertilization assay, a Dkkl1 antibody was found to significantly reduce fertilization. Human Genechips results showed that the hybridization signal intensity of DKKL1 was 405.56-fold higher in adult testis than in foetal testis. RT-PCR analysis of multiple human tissues indicated that DKKL1 mRNA was exclusively expressed in the testis. Western blot analysis also demonstrated that DKKL1 was mainly expressed in human testis with a molecular weight of approximately 34 kDa. Additionally, immunohistochemical staining showed that the DKKL1 protein was predominantly located in spermatocytes and round spermatids in human testes. An examination of the expression levels of DKKL1 in infertile male patients revealed that while no DKKL1 appeared in the testes of patients with Sertoli cell only syndrome (SCOS) or cryptorchidism, DKKL1 was observed with variable expression in patients with spermatogenic arrest.
Conclusions
These results, together with previous studies, suggest that DKKL1/Dkkl1 may play an important role in testicular development and spermatogenesis and may be an important factor in male infertility.
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作者：
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