Fluorescence properties of the Na+/H+ exchanger inhibitor HMA (5-(N,N-hexamethylene) amiloride) are modulated by intracellular pH

被引:11
作者
Giansanti, V. [1 ]
Santamaria, G. [1 ,2 ]
Torriglia, A. [3 ]
Aredia, F. [1 ]
Scovassi, A. I. [1 ]
Bottiroli, G. [1 ,2 ]
Croce, A. C. [1 ,2 ]
机构
[1] CNR, Ist Genet Mol, I-27100 Pavia, Italy
[2] Univ Pavia, Dipartimento Biol & Biotecnol, I-27100 Pavia, Italy
[3] Ctr Rech Biomed Cordeliers, INSERM, U872, Paris, France
来源
EUROPEAN JOURNAL OF HISTOCHEMISTRY | 2012年 / 56卷 / 01期
关键词
ARPE19; cells; autofluorescence; HMA; intracellular pH; fluorochrome; spectroscopy; imaging; CELLULAR-LOCALIZATION; APOPTOSIS; CELLS; PHOTOSENSITIZERS; CAMPTOTHECIN; ACCUMULATION; CARCINOMA; ACETATE;
D O I
10.4081/ejh.2012.e3
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
HMA (5-(N,N-hexamethylene)amiloride), which belongs to a family of novel amiloride derivatives, is one of the most effective inhibitors of Na+/H+ exchangers, while uneffective against Na+ channels and Na+/Ca2+ exchangers. In this study, we provided evidence that HMA can act as a fluorescent probe. In fact, human retinal ARPE19 cells incubated with HMA show an intense bluish fluorescence in the cytoplasm when observed at microscope under conventional UV-excitation conditions. Interestingly, a prolonged observation under continuous exposure to excitation lightdoes not induce great changes in cells incubated with HMA for times up to about 5 min, while an unexpected rapid increase in fluorescence signal is observed in cells incubated for longer times. The latter phenomenon is particularly evident in the perinuclear region and in discrete spots in the cytoplasm. Since HMA modulates intracellular acidity, the dependence of its fluorescence properties on medium pH and response upon irradiation have been investigated in solution, at pH 5.0 and pH 7.2. The changes in both spectral shape and amplitude emission indicate a marked pH influence on HMA fluorescence properties, making HMA exploitable as a self biomarker of pH alterations in cell studies, in the absence of perturbations induced by the administration of other exogenous dyes.
引用
收藏
页码:16 / 19
页数:4
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