Functional Involvement of Carbonic Anhydrase in the Lysosomal Response to Cadmium Exposure in Mytilus galloprovincialis Digestive Gland.
Functional Involvement of Carbonic Anhydrase in the Lysosomal Response to Cadmium Exposure in Mytilus galloprovincialis Digestive Gland.
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DOI：
10.3389/fphys.2018.00319
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发表时间：
2018
影响因子：
4
通讯作者：
Lionetto MG
中科院分区：
文献类型：
--
作者：
Caricato R;Giordano ME;Schettino T;Lionetto MG
碳酸酐酶(CA)是一种普遍存在的金属酶，在动物体内的功能包括呼吸作用、pH动态平衡、电解质转运、钙化和生物合成反应等。CA对许多物种中的痕量金属都很敏感。在贻贝中，先前的一项研究表明，镉暴露会增强消化腺中CA的活性和蛋白表达。本研究的目的是研究这种反应的功能意义，如果有的话。为此，该研究探讨了CA可能参与消化腺细胞对金属暴露的溶酶体系统反应。体内暴露于特异性CA抑制剂乙酰唑胺可显著抑制嗜酸性探针LysoSensor Green D-189的消化腺细胞溶酶体小室的酸化，在体内证实了CA对溶酶体酸化的生理贡献。在CdCl2暴露下，CA活性显著增加，与LysoSensor Green带电细胞的荧光增强平行，这反过来又表明溶酶体的增殖和/或尺寸的增大。乙酰唑胺可完全抑制镉诱导的消化腺细胞溶酶感受器荧光增强。综上所述，我们的研究结果证实了CA在紫贻贝消化腺溶酶体酸化中的作用及其参与镉暴露后溶酶体的激活。CA诱导可以对溶酶体激活过程中长时间增加的支持溶酶体酸化的H+作出生理反应。
Carbonic anhydrase (CA) is a ubiquitous metalloenzyme, whose functions in animals span from respiration to pH homeostasis, electrolyte transport, calcification, and biosynthetic reactions. CA is sensitive to trace metals in a number of species. In mussels, a previous study demonstrated CA activity and protein expression to be enhanced in digestive gland by cadmium exposure. The aim of the present work was to investigate the functional meaning, if any, of this response. To this end the study addressed the possible involvement of CA in the lysosomal system response of digestive gland cells to metal exposure. The in vivo exposure to acetazolamide, specific CA inhibitor, significantly inhibited the acidification of the lysosomal compartment in the digestive gland cells charged with the acidotropic probe LysoSensor Green D-189, demonstrating in vivo the physiological contribution of CA to the acidification of the lysosomes. Under CdCl2 exposure, CA activity significantly increased in parallel to the increase of the fluorescence of LysoSensor Green charged cells, which is in turn indicative of proliferation and/or increase in size of lysosomes. Acetazolamide exposure was able to completely inhibit the cadmium induced Lysosensor fluorescence increase in digestive gland cells. In conclusion, our results demonstrated the functional role of CA in the lysosomal acidification of Mytilus galloprovincialis digestive gland and its involvement in the lysosomal activation following cadmium exposure. CA induction could physiologically respond to a prolonged increased requirement of H+ for supporting lysosomal acidification during lysosomal activation.
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作者：
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DOI：
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