Perrotta Ida, Perri Mariarita, Santoro Marta, Panza Salvatore, Caroleo Maria C, Guido Carmela, Mete Annamaria, Cione Erika, Aquila Saveria
Departments of *Biology, Ecology and Earth Sciences (Di.B.E.S.T.) †Department of Pharmacy and Health and Nutritional Sciences, University of Calabria, Arcavacata di Rende ‡Post-graduate School in Clinical Pathology §Centro Sanitario, University of Calabria, Rende, Cosenza, Italy.
Appl Immunohistochem Mol Morphol. 2015 May-Jun;23(5):374-81. doi: 10.1097/PAI.0000000000000093.
Varicocele, an abnormal tortuosity and dilation of veins of the pampiniform plexus, is the most common identifiable and correctable cause of male infertility. It is now becoming apparent that signaling through vitamin A metabolites, such as all-trans retinoic acid (ATRA), is indispensable for spermatogenesis and disruption of retinoic acid receptor-α (RARα) function may result in male sterility and aberrant spermatogenesis. Herein, we investigated by Western blot and immunogold electron microscopy the expression profiles and subcellular localization of RARα in healthy and varicocele human sperm; in addition, we analyzed the effects of ATRA on cholesterol efflux and sperm survival utilizing enzymatic colorimetric CHOD-PAP method and Eosin Y technique, respectively. In varicocele samples, a strong reduction of RARα expression was observed. Immunogold labeling evidenced cellular location of RARα also confirming its reduced expression in "varicocele" samples. Sperm responsiveness to ATRA treatment was reduced in varicocele sperm. Our study showed that RARα is expressed in human sperm probably with a dual role in promoting both cholesterol efflux and survival. RARα might be involved in the pathogenesis of varicocele as its expression is reduced in pathologic samples. Thus, ATRA administration in procedures for artificial insemination or dietary vitamin A supplementation might represent a promising therapeutic approach for the management of male infertility.
精索静脉曲张是蔓状静脉丛静脉的异常迂曲和扩张,是男性不育最常见的可识别和可纠正原因。现在越来越明显的是,通过维生素A代谢物(如全反式维甲酸(ATRA))发出的信号对于精子发生是不可或缺的,维甲酸受体-α(RARα)功能的破坏可能导致男性不育和异常精子发生。在此,我们通过蛋白质印迹法和免疫金电子显微镜研究了RARα在健康和精索静脉曲张患者精子中的表达谱和亚细胞定位;此外,我们分别利用酶比色CHOD-PAP法和伊红Y技术分析了ATRA对胆固醇流出和精子存活的影响。在精索静脉曲张样本中,观察到RARα表达明显降低。免疫金标记证实了RARα的细胞定位,也证实了其在“精索静脉曲张”样本中的表达降低。精索静脉曲张患者的精子对ATRA治疗的反应性降低。我们的研究表明,RARα在人类精子中表达,可能在促进胆固醇流出和存活方面具有双重作用。RARα可能参与精索静脉曲张的发病机制,因为其在病理样本中的表达降低。因此,在人工授精过程中给予ATRA或通过饮食补充维生素A可能是治疗男性不育的一种有前景的治疗方法。