Légaré Christine, Thabet Michel, Sullivan Robert
Centre de Recherche en Biologie de la Reproduction, Faculté de Médecine, Université Laval, 2707 Boulevard Laurier, Sainte-Foy, Quebec, Canada G1V 4G2.
Mol Hum Reprod. 2004 Mar;10(3):197-202. doi: 10.1093/molehr/gah027. Epub 2004 Jan 29.
The Hsp70 heat-shock proteins are molecular chaperones that assist other proteins in their folding, transport and assembly into complexes. Most of these proteins are either constitutively expressed or induced by heat shock and other stresses. Heat shock proteins are required for spermatogenesis, and also protect cells from environmental hazards such as heat, radiation, and chemicals. The abdominal position of the cryptorchid testis provokes a temperature elevation which is detrimental to spermatogenesis and causes infertility. The consequences of such a stress on Hsp70 expression were evaluated in normal and cryptorchid epididymides and vas deferens. Hsp70-1 transcript from reproductive organs of normal and cryptorchid men was analysed by real-time quantitative RT-PCR, meanwhile Hsp70 protein was characterized by western blot and immunohistochemical staining analysis. Hsp70-1 mRNA and protein showed equal expression in all segments of normal epididymis and ductus deferens. Hsp70 was specific to basal cells in the epididymal epithelium. For cryptorchid patients, Hsp70-1 mRNA expression in caput epididymidis was unchanged compared with controls. However, in corpus and cauda epididymides as well as in vas deferens, the expression level of Hsp70-1 transcript was higher for the cryptorchid tissues. Changes in mRNA frequency were specifically correlated with the age of the patients. By opposition to the mRNA, western blot analysis revealed that Hsp70 protein levels were not affected by the inguinal location of the epididymis. The data show that Hsp70-1 transcript and protein were constitutively expressed in human excurrent duct and that an inguinal location can stimulate the expression of Hsp70-1 mRNA along the human epididymis and ductus deferens.
热休克蛋白70(Hsp70)是分子伴侣,可协助其他蛋白质进行折叠、运输并组装成复合物。这些蛋白质大多要么组成性表达,要么受热休克和其他应激诱导。热休克蛋白对于精子发生是必需的,还能保护细胞免受热、辐射和化学物质等环境危害。隐睾的腹部位置会引起温度升高,这对精子发生有害并导致不育。在正常和隐睾的附睾及输精管中评估了这种应激对Hsp70表达的影响。通过实时定量逆转录聚合酶链反应(RT-PCR)分析正常和隐睾男性生殖器官中的Hsp70-1转录本,同时通过蛋白质印迹法和免疫组织化学染色分析对Hsp70蛋白进行表征。Hsp70-1 mRNA和蛋白在正常附睾和输精管的所有节段中表达相等。Hsp70在附睾上皮的基底细胞中具有特异性。对于隐睾患者,附睾头中Hsp70-1 mRNA表达与对照组相比没有变化。然而,在附睾体和附睾尾以及输精管中,隐睾组织的Hsp70-1转录本表达水平更高。mRNA频率的变化与患者年龄具有特异性相关性。与mRNA相反,蛋白质印迹分析显示Hsp70蛋白水平不受附睾腹股沟位置的影响。数据表明,Hsp70-1转录本和蛋白在人类输出管道中组成性表达,腹股沟位置可刺激Hsp70-1 mRNA沿人类附睾和输精管的表达。