PMID- 34311868
OWN - NLM
STAT- MEDLINE
DCOM- 20211125
LR  - 20211125
IS  - 0091-679X (Print)
IS  - 0091-679X (Linking)
VI  - 165
DP  - 2021
TI  - Simultaneous determination of intraluminal lysosomal calcium and pH by 
      dextran-conjugated fluorescent dyes.
PG  - 199-208
LID - S0091-679X(21)00020-0 [pii]
LID - 10.1016/bs.mcb.2021.02.007 [doi]
AB  - The lysosome is the main catabolic organelle in the cell, also serving as a 
      signaling platform. Lysosomes maintain a low intraluminal pH where dozens of 
      hydrolytic enzymes degrade a wide variety of macromolecules. Besides degradation 
      of polymers, the lysosome is involved in various cellular processes, including 
      energy metabolism, plasma membrane repair and antigen presentation. Recent work 
      has shown that the lysosome is an important calcium store, modulating diverse 
      cellular functions such as membrane fusion and fission, autophagy and lysosomal 
      biogenesis. Precise measurement of free lysosomal calcium concentration has been 
      hampered by its low luminal pH, since the affinity of most calcium probes 
      decreases with higher proton concentration. Here we detailed an adapted protocol 
      for the simultaneous measurement of lysosomal pH and calcium using 
      dextran-conjugated ratiometric fluorescent dyes. As compared with indirect 
      measurements of lysosomal calcium release using genetically-encoded calcium 
      indicators (GECIs), the present method offers the possibility of obtaining 
      pH-corrected, intraluminal calcium concentrations at single lysosome resolution. 
      It also enables simultaneous temporal resolution of lysosomal calcium and pH.
CI  - Copyright (c) 2021 Elsevier Inc. All rights reserved.
FAU - Pihan, Philippe
AU  - Pihan P
AD  - Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, 
      Santiago, Chile; Center for Geroscience, Brain Health and Metabolism (GERO), 
      Santiago, Chile; Program of Cellular and Molecular Biology, Institute of 
      Biomedical Sciences, University of Chile, Santiago, Chile.
FAU - Nunes-Hasler, Paula
AU  - Nunes-Hasler P
AD  - Department of Cell Physiology and Metabolism, University of Geneva, Geneva, 
      Switzerland; Department of Pathology and Immunology, University of Geneva, 
      Geneva, Switzerland.
FAU - Demaurex, Nicolas
AU  - Demaurex N
AD  - Department of Cell Physiology and Metabolism, University of Geneva, Geneva, 
      Switzerland; Department of Pathology and Immunology, University of Geneva, 
      Geneva, Switzerland. Electronic address: Nicolas.Demaurex@unige.ch.
FAU - Hetz, Claudio
AU  - Hetz C
AD  - Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, 
      Santiago, Chile; Center for Geroscience, Brain Health and Metabolism (GERO), 
      Santiago, Chile; Program of Cellular and Molecular Biology, Institute of 
      Biomedical Sciences, University of Chile, Santiago, Chile; Buck Institute for 
      Research on Aging, Novato, CA, United States. Electronic address: 
      chetz@uchile.cl.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20210319
PL  - United States
TA  - Methods Cell Biol
JT  - Methods in cell biology
JID - 0373334
RN  - 0 (Dextrans)
RN  - 0 (Fluorescent Dyes)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - *Calcium
MH  - Dextrans
MH  - *Fluorescent Dyes
MH  - Hydrogen-Ion Concentration
MH  - Lysosomes
OTO - NOTNLM
OT  - Confocal microscopy
OT  - Lysosomal calcium
OT  - Lysosomal signaling
OT  - Ratiometric fluorescent probes
EDAT- 2021/07/28 06:00
MHDA- 2021/11/26 06:00
CRDT- 2021/07/27 05:47
PHST- 2021/07/27 05:47 [entrez]
PHST- 2021/07/28 06:00 [pubmed]
PHST- 2021/11/26 06:00 [medline]
AID - S0091-679X(21)00020-0 [pii]
AID - 10.1016/bs.mcb.2021.02.007 [doi]
PST - ppublish
SO  - Methods Cell Biol. 2021;165:199-208. doi: 10.1016/bs.mcb.2021.02.007. Epub 2021 
      Mar 19.