PMID- 36427748
OWN - NLM
STAT- MEDLINE
DCOM- 20230109
LR  - 20230114
IS  - 1873-4596 (Electronic)
IS  - 0891-5849 (Linking)
VI  - 194
DP  - 2023 Jan
TI  - Real-time simultaneous imaging of temporal alterations in cytoplasmic and 
      mitochondrial redox in single cells during cell division and cell death.
PG  - 33-41
LID - S0891-5849(22)00994-7 [pii]
LID - 10.1016/j.freeradbiomed.2022.11.031 [doi]
AB  - Cytosolic and organelle redox are highly interrelated, and their alterations play 
      critical roles in both physiological and pathological cell states. This highly 
      regulated process is crucial in life-death decisions of cells. Among organelles, 
      the mitochondrion is the major source of intracellular-ROS and contributes to 
      oxidation damage-induced cell death. Increase in cytosolic-redox and 
      mitochondrial-redox is evident in cells undergoing diverse forms of cell death, 
      such as apoptosis, necrosis, and necroptosis. The hierarchical profiling of redox 
      signaling at the cytosol and mitochondria in a single cell is important to 
      understand the relative contribution of each species in the initiation and 
      shaping of cell death. Here, we demonstrate the potential application of 
      ratiometric redox GFP (roGFP) and intensity-based redox-sensitive RFP (rxRFP) 
      targeted to mitochondria in revealing both rapid and slow progressing changes in 
      redox during cell division and in cells undergoing multiple modes of cell death. 
      To generate imaging quality signal, single-cell clones stably expressing both 
      roGFP at the cytosol and rxRFP probes targeted to mitochondria were generated. 
      The cells provided sufficient temporal resolution with imaging-ready signal for 
      the real-time visualization of rapidly progressing redox alterations at the 
      cytosol and mitochondria. The long-time imaging of the cells revealed that a 
      moderate increase in cytosolic ROS marks the division stage. Similarly, distinct 
      forms of cell death trigger a unique and temporally regulated redox change at the 
      cytosol and mitochondria, suggesting the potential utility of the sensor cells to 
      dissect the nature of cell death pathways induced by specific forms of stress.
CI  - Copyright (c) 2022 The Authors. Published by Elsevier Inc. All rights reserved.
FAU - Chandrasekharan, Aneesh
AU  - Chandrasekharan A
AD  - Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, 
      Thycaud P.O., Thiruvananthapuram, Kerala, 695014, India.
FAU - Varadarajan, Shankara Narayanan
AU  - Varadarajan SN
AD  - Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, 
      Thycaud P.O., Thiruvananthapuram, Kerala, 695014, India.
FAU - Lekshmi, Asha
AU  - Lekshmi A
AD  - Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, 
      Thycaud P.O., Thiruvananthapuram, Kerala, 695014, India.
FAU - Santhoshkumar, T R
AU  - Santhoshkumar TR
AD  - Cancer Research Program, Rajiv Gandhi Centre for Biotechnology, Poojappura, 
      Thycaud P.O., Thiruvananthapuram, Kerala, 695014, India. Electronic address: 
      trsanthosh@rgcb.res.in.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221123
PL  - United States
TA  - Free Radic Biol Med
JT  - Free radical biology & medicine
JID - 8709159
RN  - 0 (Reactive Oxygen Species)
SB  - IM
MH  - Cytosol/metabolism
MH  - Reactive Oxygen Species/metabolism
MH  - *Mitochondria/metabolism
MH  - Oxidation-Reduction
MH  - Cell Death
MH  - Cell Division
OTO - NOTNLM
OT  - Cell death
OT  - Cytosolic redox
OT  - Fluorescent probes
OT  - Mitochondrial redox
OT  - Redox imaging
COIS- Declaration of competing interest The authors declare that they have no competing 
      interests.
EDAT- 2022/11/26 06:00
MHDA- 2023/01/10 06:00
CRDT- 2022/11/25 19:26
PHST- 2022/11/03 00:00 [received]
PHST- 2022/11/17 00:00 [accepted]
PHST- 2022/11/26 06:00 [pubmed]
PHST- 2023/01/10 06:00 [medline]
PHST- 2022/11/25 19:26 [entrez]
AID - S0891-5849(22)00994-7 [pii]
AID - 10.1016/j.freeradbiomed.2022.11.031 [doi]
PST - ppublish
SO  - Free Radic Biol Med. 2023 Jan;194:33-41. doi: 
      10.1016/j.freeradbiomed.2022.11.031. Epub 2022 Nov 23.