Caceres Ana I, Obeso Ana, Gonzalez Constancio, Rocher Asuncion
Departamento de Bioquímica, Biología Molecular y Fisiología, Facultad de Medicina/Instituto de Biología y Genética Molecular (IBGM), Universidad de Valladolid/CSIC, Valladolid, Spain.
J Neurochem. 2007 Jul;102(1):231-45. doi: 10.1111/j.1471-4159.2007.04465.x.
We have assessed the expression, molecular identification and functional role of Na+ channels (Na(v)) in carotid bodies (CB) obtained from normoxic and chronically hypoxic adult rats. Veratridine evoked release of catecholamines (CA) from an in vitro preparation of intact CBs obtained from normoxic animals, the response being Ca2+ and Na+-dependent and sensitive to tetrodotoxin (TTX). TTX inhibited by 25-50% the CA release response evoked by graded hypoxia. Immunoblot assays demonstrated the presence of Na(v)alpha-subunit (c. 220 kDa) in crude homogenates from rat CBs, being evident an up-regulation (60%) of this protein in the CBs obtained from chronically hypoxic rats (10% O2; 7 days). This up-regulation was accompanied by an enhanced TTX-sensitive release response to veratridine, and by an enhanced ventilatory response to acute hypoxic stimuli. RT-PCR studies demonstrated the expression of mRNA for Na(v)1.1, Na(v)1.2, Na(v)1.3, Na(v)1.6 and Na(v)1.7 isoforms. At least three isoforms, Na(v)1.1, Na(v)1.3 and Na(v)1.6 co-localized with tyrosine hydroxylase in all chemoreceptor cells. RT-PCR and immunocytochemistry indicated that Na(v)1.1 isoform was up-regulated by chronic hypoxia in chemoreceptor cells. We conclude that Na(v) up-regulation represents an adaptive mechanism to increase chemoreceptor sensitivity during acclimatization to sustained hypoxia as evidenced by enhanced ventilatory responses to acute hypoxic tests.
我们评估了从常氧和慢性低氧成年大鼠获取的颈动脉体(CB)中钠通道(Na(v))的表达、分子鉴定及功能作用。藜芦碱可诱发从常氧动物完整CB的体外制备物中释放儿茶酚胺(CA),该反应依赖于Ca2+和Na+,且对河豚毒素(TTX)敏感。TTX可抑制分级低氧诱发的CA释放反应的25 - 50%。免疫印迹分析表明,大鼠CB的粗匀浆中存在Na(v)α亚基(约220 kDa),在慢性低氧大鼠(10% O2;7天)获取的CB中该蛋白明显上调(60%)。这种上调伴随着对藜芦碱的TTX敏感释放反应增强,以及对急性低氧刺激的通气反应增强。逆转录聚合酶链反应(RT-PCR)研究表明,Na(v)1.1、Na(v)1.2、Na(v)1.3、Na(v)1.6和Na(v)1.7亚型的mRNA有表达。至少三种亚型,即Na(v)1.1、Na(v)1.3和Na(v)1.6在所有化学感受细胞中与酪氨酸羟化酶共定位。RT-PCR和免疫细胞化学表明,化学感受细胞中Na(v)1.1亚型在慢性低氧时上调。我们得出结论,Na(v)上调代表一种适应性机制,可在适应持续性低氧过程中增加化学感受器敏感性,急性低氧试验中增强的通气反应证明了这一点。