PMID- 26344758
OWN - NLM
STAT- MEDLINE
DCOM- 20160915
LR  - 20240324
IS  - 1095-9130 (Electronic)
IS  - 1046-2023 (Print)
IS  - 1046-2023 (Linking)
VI  - 90
DP  - 2015 Nov 15
TI  - Use of genetically-encoded calcium indicators for live cell calcium imaging and 
      localization in virus-infected cells.
PG  - 28-38
LID - S1046-2023(15)30075-X [pii]
LID - 10.1016/j.ymeth.2015.09.004 [doi]
AB  - Calcium signaling is a ubiquitous and versatile process involved in nearly every 
      cellular process, and exploitation of host calcium signals is a common strategy 
      used by viruses to facilitate replication and cause disease. Small molecule 
      fluorescent calcium dyes have been used by many to examine changes in host cell 
      calcium signaling and calcium channel activation during virus infections, but 
      disadvantages of these dyes, including poor loading and poor long-term retention, 
      complicate analysis of calcium imaging in virus-infected cells due to changes in 
      cell physiology and membrane integrity. The recent expansion of 
      genetically-encoded calcium indicators (GECIs), including blue and red-shifted 
      color variants and variants with calcium affinities appropriate for calcium 
      storage organelles like the endoplasmic reticulum (ER), make the use of GECIs an 
      attractive alternative for calcium imaging in the context of virus infections. 
      Here we describe the development and testing of cell lines stably expressing both 
      green cytoplasmic (GCaMP5G and GCaMP6s) and red ER-targeted (RCEPIAer) GECIs. 
      Using three viruses (rotavirus, poliovirus and respiratory syncytial virus) 
      previously shown to disrupt host calcium homeostasis, we show the GECI cell lines 
      can be used to detect simultaneous cytoplasmic and ER calcium signals. Further, 
      we demonstrate the GECI expression has sufficient stability to enable long-term 
      confocal imaging of both cytoplasmic and ER calcium during the course of virus 
      infections.
CI  - Copyright (c) 2015 Elsevier Inc. All rights reserved.
FAU - Perry, Jacob L
AU  - Perry JL
AD  - Department of Molecular Virology and Microbiology and Alkek Center for 
      Metagenomic and Microbiome Research, Baylor College of Medicine, Houston, TX 
      77030, United States.
FAU - Ramachandran, Nina K
AU  - Ramachandran NK
AD  - Department of Molecular Virology and Microbiology and Alkek Center for 
      Metagenomic and Microbiome Research, Baylor College of Medicine, Houston, TX 
      77030, United States.
FAU - Utama, Budi
AU  - Utama B
AD  - Shared Equipment Authority, Rice University, Houston, TX 77030, United States.
FAU - Hyser, Joseph M
AU  - Hyser JM
AD  - Department of Molecular Virology and Microbiology and Alkek Center for 
      Metagenomic and Microbiome Research, Baylor College of Medicine, Houston, TX 
      77030, United States. Electronic address: joseph.hyser@bcm.edu.
LA  - eng
GR  - P30 AI027767/AI/NIAID NIH HHS/United States
GR  - CA125123/CA/NCI NIH HHS/United States
GR  - AI036211/AI/NIAID NIH HHS/United States
GR  - K01 DK093657/DK/NIDDK NIH HHS/United States
GR  - K01DK093657/DK/NIDDK NIH HHS/United States
GR  - P30 CA125123/CA/NCI NIH HHS/United States
GR  - P30 AI036211/AI/NIAID NIH HHS/United States
GR  - S10 RR024574/RR/NCRR NIH HHS/United States
GR  - RR024574/RR/NCRR NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20150903
PL  - United States
TA  - Methods
JT  - Methods (San Diego, Calif.)
JID - 9426302
RN  - 0 (Indicators and Reagents)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Calcium/*analysis
MH  - *Calcium Signaling
MH  - Cell Line
MH  - Chlorocebus aethiops
MH  - Endoplasmic Reticulum/metabolism
MH  - HEK293 Cells
MH  - *Host-Pathogen Interactions
MH  - Humans
MH  - Indicators and Reagents/chemistry
MH  - Microscopy, Fluorescence/*methods
MH  - Poliovirus/metabolism
MH  - Respiratory Syncytial Viruses/metabolism
MH  - Rotavirus/metabolism
PMC - PMC4655165
MID - NIHMS726459
OTO - NOTNLM
OT  - Endoplasmic reticulum
OT  - Enterovirus
OT  - GCaMP5G
OT  - GCaMP6s
OT  - RCEPIAer
OT  - Respiratory syncytial virus
OT  - Rotavirus
EDAT- 2015/09/08 06:00
MHDA- 2016/09/16 06:00
PMCR- 2016/11/15
CRDT- 2015/09/08 06:00
PHST- 2015/05/16 00:00 [received]
PHST- 2015/08/31 00:00 [revised]
PHST- 2015/09/01 00:00 [accepted]
PHST- 2015/09/08 06:00 [entrez]
PHST- 2015/09/08 06:00 [pubmed]
PHST- 2016/09/16 06:00 [medline]
PHST- 2016/11/15 00:00 [pmc-release]
AID - S1046-2023(15)30075-X [pii]
AID - 10.1016/j.ymeth.2015.09.004 [doi]
PST - ppublish
SO  - Methods. 2015 Nov 15;90:28-38. doi: 10.1016/j.ymeth.2015.09.004. Epub 2015 Sep 3.