PMID- 35237413
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20221222
IS  - 2045-452X (Print)
IS  - 2045-4538 (Electronic)
IS  - 2045-452X (Linking)
VI  - 11
IP  - 1
DP  - 2022 Feb
TI  - JC-10 probe as a novel method for analyzing the mitochondrial membrane potential 
      and cell stress in whole zebrafish embryos.
PG  - 77-87
LID - 10.1093/toxres/tfab114 [doi]
AB  - BACKGROUND: A sensitive method to investigate cellular stress and cytotoxicity is 
      based on measuring mitochondrial membrane potential. Recently, JC-10, was 
      developed to measure mitochondrial membrane potential in vitro and used as an 
      indicator for cytotoxicity. Yet, JC-10 has never been used in vivo (whole 
      organism). In normal cells, JC-10 concentrates in the mitochondrial matrix, where 
      it forms red fluorescent aggregates. However, in apoptotic/necrotic cells, JC-10 
      diffuses out of the mitochondria, changes to monomeric form, and stains cells in 
      green. Here, we aimed to develop and optimize a JC-10 assay to measure 
      cytotoxicity in zebrafish embryo. We also investigated the effectiveness of JC-10 
      assay by comparing it to common cytotoxicity assays. METHODS: Zebrafish embryos 
      were exposed to a toxic surfactant AEO-7 at no observed effect concentration 
      (6.4 mug/L), and then cytotoxicity was measured using (i) JC-10 mitochondrial 
      assay, (ii) acridine orange (AO), (iii) TUNEL assay, and (iv) measuring the level 
      of Hsp70 by western blotting. RESULTS: As compared to the negative control, 
      embryos treated with NOEC of AEO-7 did not show significant cytotoxicity when 
      assessed by AO, TUNEL or western blotting. However, when JC-10 was used under the 
      same experimental conditions, a significant increase of green:red fluorescent 
      ratio signal was detected in the AEO-7 treated embryos, indicating mitochondrial 
      damage and cellular cytotoxicity. Noteworthy, the observed green: red ratio 
      increase was dose dependent, suggesting specificity of the JC-10 assay. 
      CONCLUSION: JC-10 is a sensitive in vivo method, thus, can be used as surrogate 
      assay to measure cytotoxicity in whole zebrafish embryos.
CI  - (c) The Author(s) 2021. Published by Oxford University Press. All rights reserved. 
      For Permissions, please email: journals.permissions@oup.com.
FAU - Younes, Nadin
AU  - Younes N
AD  - Biomedical Research Center, QU Health, Qatar University, P.O. Box 2713 Doha, 
      Qatar.
FAU - Alsahan, Bana S
AU  - Alsahan BS
AD  - Department of Biomedical Science, College of Health Sciences, QU Health, Qatar 
      University, P.O. Box 2713 Doha, Qatar.
FAU - Al-Mesaifri, Asmaa J
AU  - Al-Mesaifri AJ
AD  - Department of Biomedical Science, College of Health Sciences, QU Health, Qatar 
      University, P.O. Box 2713 Doha, Qatar.
FAU - Da'as, Sahar I
AU  - Da'as SI
AD  - Department of Human Genetics, Sidra Medicine, P.O. Box 26999, Doha, Qatar.
FAU - Pintus, Gianfranco
AU  - Pintus G
AD  - Department of Medical Laboratory Sciences, University of Sharjah, P.O. Box 27272 
      Sharjah, United Arab Emirates.
FAU - Majdalawieh, Amin F
AU  - Majdalawieh AF
AD  - Department of Biology, Chemistry, and Environmental Sciences, College of Arts and 
      Sciences, American University of Sharjah, P.O. Box 26666 Sharjah, United Arab 
      Emirates.
FAU - Nasrallah, Gheyath K
AU  - Nasrallah GK
AUID- ORCID: 0000-0001-9252-1038
AD  - Biomedical Research Center, QU Health, Qatar University, P.O. Box 2713 Doha, 
      Qatar.
LA  - eng
PT  - Journal Article
DEP - 20211221
PL  - England
TA  - Toxicol Res (Camb)
JT  - Toxicology research
JID - 101587950
PMC - PMC8882781
OTO - NOTNLM
OT  - AEO-7
OT  - JC-10 probe
OT  - cytotoxicity
OT  - mitochondria
OT  - zebrafish
EDAT- 2022/03/04 06:00
MHDA- 2022/03/04 06:01
PMCR- 2022/12/21
CRDT- 2022/03/03 05:35
PHST- 2021/01/22 00:00 [received]
PHST- 2021/10/22 00:00 [revised]
PHST- 2021/11/03 00:00 [accepted]
PHST- 2022/03/03 05:35 [entrez]
PHST- 2022/03/04 06:00 [pubmed]
PHST- 2022/03/04 06:01 [medline]
PHST- 2022/12/21 00:00 [pmc-release]
AID - tfab114 [pii]
AID - 10.1093/toxres/tfab114 [doi]
PST - epublish
SO  - Toxicol Res (Camb). 2021 Dec 21;11(1):77-87. doi: 10.1093/toxres/tfab114. 
      eCollection 2022 Feb.