PMID- 31758454
OWN - NLM
STAT- MEDLINE
DCOM- 20210111
LR  - 20210111
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Print)
IS  - 1064-3745 (Linking)
VI  - 2071
DP  - 2020
TI  - Genetic Indicators for Calcium Signaling Studies in Toxoplasma gondii.
PG  - 187-207
LID - 10.1007/978-1-4939-9857-9_11 [doi]
AB  - Fluctuations of the cytosolic calcium ion (Ca(2+)) concentration regulate a 
      variety of cellular functions in all eukaryotes. Cells express a sophisticated 
      set of mechanisms to balance the cytosolic Ca(2+) levels and the signals that 
      elevate Ca(2+) in the cytosol are compensated by mechanisms that reduce it. 
      Alterations in Ca(2+)-dependent homeostatic mechanisms are the cause of many 
      prominent diseases in humans, such as heart failure or neuronal death.The genetic 
      tractability of Toxoplasma gondii and the availability of genetic tools enabled 
      the use of Genetically Encoded Calcium Indicators (GECIs) expressed in the 
      cytoplasm, which started a new era in the studies of Toxoplasma calcium 
      signaling. It was finally possible to see Ca(2+) oscillations prior to exit of 
      the parasite from host cells. Years after Endo et al showed that ionophores 
      triggered egress, the assumption that oscillations occur prior to egress from 
      host cells has been validated by experiments using GECIs. GECIs allowed the 
      visualization of specific Ca(2+) signals in live intracellular parasites and to 
      distinguish these signals from host cell calcium fluctuations. In this chapter we 
      present an overview describing "tried and true" methods of our lab who pioneered 
      the first use of GECI's in Toxoplasma, including GECI choice, methodology for 
      transfection and selection of ideal clones, their characterization, and the use 
      of GECI-expressing parasites for fluorometric and microscopic analysis.
FAU - Vella, Stephen A
AU  - Vella SA
AD  - Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, 
      GA, USA.
FAU - Calixto, Abigail
AU  - Calixto A
AD  - Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, 
      GA, USA.
FAU - Asady, Beejan
AU  - Asady B
AD  - Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, 
      GA, USA.
FAU - Li, Zhu-Hong
AU  - Li ZH
AD  - Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, 
      GA, USA.
FAU - Moreno, Silvia N J
AU  - Moreno SNJ
AD  - Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, 
      GA, USA. smoreno@uga.edu.
AD  - Department of Cellular Biology, University of Georgia, Athens, GA, USA. 
      smoreno@uga.edu.
LA  - eng
GR  - R01 AI096836/AI/NIAID NIH HHS/United States
GR  - R01 AI128356/AI/NIAID NIH HHS/United States
GR  - R21 AI110027/AI/NIAID NIH HHS/United States
GR  - T32 AI060546/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Ionophores)
RN  - 0 (Protozoan Proteins)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - Calcium/metabolism
MH  - Calcium Signaling/physiology
MH  - Cytoplasm/metabolism
MH  - Cytosol/metabolism
MH  - Host-Parasite Interactions
MH  - Humans
MH  - Ionophores/metabolism
MH  - Protozoan Proteins/metabolism
MH  - Toxoplasma/*metabolism
PMC - PMC7294879
MID - NIHMS1594056
OTO - NOTNLM
OT  - Calcium signaling
OT  - Egress
OT  - Fura2 measurements
OT  - GCaMPs
OT  - GECOs
OT  - Genetic indicators
OT  - Time lapse
OT  - Toxoplasma
EDAT- 2019/11/24 06:00
MHDA- 2021/01/12 06:00
PMCR- 2020/06/15
CRDT- 2019/11/24 06:00
PHST- 2019/11/24 06:00 [entrez]
PHST- 2019/11/24 06:00 [pubmed]
PHST- 2021/01/12 06:00 [medline]
PHST- 2020/06/15 00:00 [pmc-release]
AID - 10.1007/978-1-4939-9857-9_11 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2020;2071:187-207. doi: 10.1007/978-1-4939-9857-9_11.