Is cadmium chloride-induced inter-sertoli tight junction permeability barrier disruption a suitable in vitro model to study the events of junction disassembly during spermatogenesis in the rat testis?
Is cadmium chloride-induced inter-sertoli tight junction permeability barrier disruption a suitable in vitro model to study the events of junction disassembly during spermatogenesis in the rat testis?
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DOI：
10.1210/en.142.5.1878
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发表时间：
2001-05-01
期刊：
影响因子：
4.8
通讯作者：
Cheng, CY
中科院分区：
文献类型：
--
作者：
Chung, NPY;Cheng, CY
精子发生过程中生殖细胞运动的事件是由连接的拆卸和重组的间歇性阶段。尽管体外培养的原代支持细胞可用于研究连接重组，但仍缺乏研究连接解体事件的体外模型。我们已经评估了CdCl 2诱导的支持细胞间紧密连接（TJ）渗透性屏障在体外破坏是否可以填补这一空白。当支持细胞（1.2 × 10（6）细胞/cm（2））在Matrigel包被的双室单位上培养以允许支持间TJ组装时，支持细胞上皮的跨上皮电阻稳定上升。将这些细胞在第1天（即分离后24小时）暴露于5-10 μ M的CdCl 2 8小时，可以剂量依赖性地干扰Sertoli TJ间组装，而没有任何明显的细胞毒性。同样地，当细胞在第4天暴露于CdCl 2（0.1-5 μ M）8小时后，间-Sertoli TJ已经组装，CdCl 2也扰乱间-Sertoli TJ渗透屏障的维持剂量依赖性，没有细胞毒性的迹象。尽管即使在去除CdCl 2后，扰动的支持细胞间TJ也不能重新密封，但在去除CdCl 2后，1 × 10（-9）M睾酮（T）的存在允许支持细胞间TJ屏障重新密封，而2 × 10（-7）M睾酮的存在甚至可以保护支持细胞免受CdCl 2诱导的损伤。更重要的是，重新组装后CdCl 2诱导的TJ中断间Sertoli TJ伴随着细胞基因表达的变化occludin和尿激酶纤溶酶原激活剂，模仿他们的模式在体外组装间Sertoli TJ没有CdCl 2处理。基于这些结果，很明显，CdCl 2诱导的支持间TJ拆卸是一个潜在的体外模型，以研究连接拆卸的事件。
The events of germ cell movement during spermatogenesis are composed of intermittent phases of junction disassembly and reassembly. Although primary Sertoli cells cultured in vitro can be used to study junction reassembly, an in vitro model to study the events of junction disassembly is still lacking. We have assessed whether the CdCl2-induced inter-Sertoli tight junction (TJ) permeability barrier disruption in vitro can fill this gap. When Sertoli cells (1.2 X 10(6) cells/cm(2)) were cultured on Matrigel-coated bicameral units to allow the assembly of inter-Sertoli TJs, it was manifested by a steady rise in transepithelial electrical resistance across the Sertoli cell epithelia. Exposure of these cells on day 1 (i.e. 24h after their isolation) to CdCl2, at 5-10 muM for 8 h could perturb the inter-Sertoli TJ assembly dose dependently without any apparent cytotoxicity. Likewise, when cells were exposed to CdCl2 (0.1-5 muM) on day 4 for 8 h after inter-Sertoli TJs were already assembled, CdCl2 also perturbed the maintenance of inter-Sertoli TJ permeability barrier dose dependently without signs of cell cytotoxicity. Although the perturbed inter-Sertoli TJs were not capable of resealing even after the removal of CdCl2, the presence of testosterone (T) at 1 x 10(-9) M allowed resealing of the inter-Sertoli TJ barrier after CdCl2 was removed, whereas the presence of 2 x 10(-7) M testosterone even protected Sertoli cells from CdCl2-induced damage. More important, the reassembly of inter-Sertoli TJs after CdCl2-induced TJ disruption was accompanied by changes in cellular gene expression of occludin and urokinase plasminogen activator, which mimicked their patterns during inter-Sertoli TJ assembly in vitro without CdCl2 treatment. Based on these results, it is apparent that CdCl2-induced inter-Sertoli TJ disassembly is a potential in vitro model to study the events of junction disassembly.