Quintas Luis Eduardo M, Cunha Valéria M N, Scaramello Christianne B V, da Silva Cláudia L M, Caricati-Neto Afonso, Lafayette Simone S L, Jurkiewicz Aron, Noël François
Departamento de Farmacologia Básica e Clínica, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Cidade Universitária, 21941-590, Rio de Janeiro, Brazil.
Eur J Pharmacol. 2005 Nov 21;525(1-3):54-9. doi: 10.1016/j.ejphar.2005.10.006. Epub 2005 Nov 14.
The activity and protein expression of plasma membrane and sarco(endo)plasmic reticulum (Ca2+-Mg2+)ATPases and ryanodine receptors were investigated in surgically denervated rat vas deferens. The function of thapsigargin-sensitive but not thapsigargin-resistant (Ca2+-Mg2+)ATPase (from sarco(endo)plasmic reticulum and plasma membrane, respectively), evidenced by enzyme activity and Ca2+ uptake experiments, was significantly depressed by 30-50% when compared to innervated vas. Western blots showed that such reduction in sarco(endo)plasmic reticulum (Ca2+-Mg2+)ATPase performance was accompanied by a decrement of similar magnitude in sarco(endo)plasmic reticulum (Ca2+-Mg2+)ATPase type 2 protein expression, without any significant change in plasma membrane (Ca2+-Mg2+)ATPase expression. Finally, [3H]ryanodine binding revealed that the density of ryanodine binding sites was reduced by 45% after denervation without modification in affinity. The present findings demonstrate that sarco(endo)plasmic reticulum proteins involved in intracellular calcium homeostasis are clearly down-regulated and brings further evidence of a modified calcium translocation in denervated rat vas deferens.
在手术去神经支配的大鼠输精管中,研究了质膜和肌浆(内质)网(Ca2+-Mg2+)ATP酶及兰尼碱受体的活性和蛋白表达。毒胡萝卜素敏感型(而非毒胡萝卜素抵抗型)(Ca2+-Mg2+)ATP酶(分别来自肌浆(内质)网和质膜)的功能,通过酶活性和Ca2+摄取实验证明,与神经支配的输精管相比,显著降低了30 - 50%。蛋白质免疫印迹显示,肌浆(内质)网(Ca2+-Mg2+)ATP酶性能的这种降低伴随着肌浆(内质)网(Ca2+-Mg2+)2型ATP酶蛋白表达类似程度的减少,而质膜(Ca2+-Mg2+)ATP酶表达无任何显著变化。最后,[3H]兰尼碱结合显示,去神经支配后兰尼碱结合位点的密度降低了45%,亲和力无改变。本研究结果表明,参与细胞内钙稳态的肌浆(内质)网蛋白明显下调,并为去神经支配的大鼠输精管中钙转运改变提供了进一步证据。