Ronco Virginia, Potenza Duilio Michele, Denti Federico, Vullo Sabrina, Gagliano Giuseppe, Tognolina Marialuisa, Guerra Germano, Pinton Paolo, Genazzani Armando A, Mapelli Lisa, Lim Dmitry, Moccia Francesco
Department of Pharmaceutical Sciences, Università del Piemonte Orientale "Amedeo Avogadro", 28100 Novara, Italy.
Laboratory of General Physiology, Department of Biology and Biotechnology "Lazzaro Spallanzani", University of Pavia, 27100 Pavia, Italy.
Cell Calcium. 2015 Feb;57(2):89-100. doi: 10.1016/j.ceca.2015.01.001. Epub 2015 Jan 21.
Nicotinic acid adenine dinucleotide phosphate (NAADP) serves as the ideal trigger of spatio-temporally complex intracellular Ca(2+) signals. However, the identity of the intracellular Ca(2+) store(s) recruited by NAADP, which may include either the endolysosomal (EL) or the endoplasmic reticulum (ER) Ca(2+) pools, is still elusive. Here, we show that the Ca(2+) response to NAADP was suppressed by interfering with either EL or ER Ca(2+) sequestration. The measurement of EL and ER Ca(2+) levels by using selectively targeted aequorin unveiled that the preventing ER Ca(2+) storage also affected ER Ca(2+) loading and vice versa. This indicates that a functional Ca(2+)-mediated cross-talk exists at the EL-ER interface and exerts profound implications for the study of NAADP-induced Ca(2+) signals. Extreme caution is warranted when dissecting NAADP targets by pharmacologically inhibiting EL and/or the ER Ca(2+) pools. Moreover, Ca(2+) transfer between these compartments might be essential to regulate vital Ca(2+)-dependent processes in both organelles.
烟酰胺腺嘌呤二核苷酸磷酸(NAADP)是时空复杂的细胞内Ca(2+)信号的理想触发因子。然而,NAADP募集的细胞内Ca(2+)储存库(可能包括内溶酶体(EL)或内质网(ER)Ca(2+)池)的身份仍然难以捉摸。在这里,我们表明,通过干扰EL或ER Ca(2+)螯合,对NAADP的Ca(2+)反应受到抑制。使用选择性靶向水母发光蛋白测量EL和ER Ca(2+)水平发现,阻止ER Ca(2+)储存也会影响ER Ca(2+)负载,反之亦然。这表明在EL-ER界面存在功能性的Ca(2+)介导的串扰,对NAADP诱导的Ca(2+)信号的研究具有深远意义。在通过药理抑制EL和/或ER Ca(2+)池来剖析NAADP靶点时,必须格外谨慎。此外,这些区室之间的Ca(2+)转移对于调节两个细胞器中重要的Ca(2+)依赖性过程可能至关重要。