Carlin Ryan W, Lee Jun Ho, Marcus Daniel C, Schultz Bruce D
Department of Anatomy and Physiology, Kansas State University, Manhattan, Kansas 66502-5802, USA.
Biol Reprod. 2003 Mar;68(3):1027-34. doi: 10.1095/biolreprod.102.009381.
Experiments were conducted to determine the responsiveness of human vas deferens epithelial cell monolayers to adenosine and related agonists. Human abdominal vas deferens epithelial cells have been isolated from adult tissues and grown to confluence on permeable supports. All cells exhibit intense ZO-1 and cytokeratin immunoreactivity. Cultured cell monolayers exhibit high electrical resistance with a lumen-negative potential difference and short circuit current (I(sc)) indicative of anion secretion and/or cation absorption. A portion of the basal I(sc) is inhibited by amiloride. Amiloride-sensitive I(sc) is enhanced by exposure to glucocorticoids and is Na(+) dependent, indicating the presence of epithelial sodium channel-mediated Na(+) absorption. Epithelial anion secretion and intracellular generation of cAMP are acutely stimulated by adenosine and the adenosine receptor agonist 5'-(N-ethylcarboxamido)adenosine (NECA), with these effects being fully blocked by 8-phenyltheophylline. Adenosine receptors are localized to the apical membrane of the epithelial cells, as basolateral adenosine is without effect. Freshly excised human vas deferens recapitulate observations made on cultured epithelia when evaluated with the self-referencing vibrating probe: amiloride inhibition of basal ion transport, stimulation by adenosine, and inhibition by 8-phenyltheophyline. These results demonstrate that adult human vas deferens epithelium actively transports ions to generate the luminal environment of the deferent duct. Thus, vas deferens epithelium likely plays an active role in male fertility, and interventions that modulate epithelial function might be exploited to treat male-factor infertility or in contraception.
进行实验以确定人输精管上皮细胞单层对腺苷及相关激动剂的反应性。人腹段输精管上皮细胞已从成人组织中分离出来，并在可渗透支持物上生长至汇合。所有细胞均表现出强烈的ZO-1和细胞角蛋白免疫反应性。培养的细胞单层表现出高电阻，具有管腔负电位差和短路电流（I(sc)），表明存在阴离子分泌和/或阳离子吸收。基础I(sc)的一部分被氨氯吡咪抑制。氨氯吡咪敏感的I(sc)在暴露于糖皮质激素时增强，且依赖于Na(+)，表明存在上皮钠通道介导的Na(+)吸收。腺苷和腺苷受体激动剂5'-(N-乙基甲酰胺基)腺苷（NECA）可急性刺激上皮阴离子分泌和细胞内cAMP的产生，这些作用被8-苯基茶碱完全阻断。腺苷受体定位于上皮细胞的顶端膜，因为基底外侧的腺苷没有作用。当用自参考振动探针评估时，新鲜切除的人输精管重现了在培养上皮细胞上的观察结果：氨氯吡咪对基础离子转运的抑制、腺苷的刺激以及8-苯基茶碱的抑制。这些结果表明，成人输精管上皮细胞主动转运离子以产生输精管的管腔环境。因此，输精管上皮细胞可能在男性生育中发挥积极作用，调节上皮功能的干预措施可能被用于治疗男性因素不孕症或用于避孕。
