PMID- 35960624
OWN - NLM
STAT- MEDLINE
DCOM- 20221103
LR  - 20221222
IS  - 1559-6095 (Electronic)
IS  - 1559-6095 (Linking)
VI  - 2022
IP  - 11
DP  - 2022 Nov 1
TI  - Calcium Imaging of Anopheles coluzzii Mosquito Antennae Expressing the Calcium 
      Indicator GCaMP6f.
PG  - Pdb.prot107918
LID - 10.1101/pdb.prot107918 [doi]
AB  - Calcium imaging is a technique used to measure functional neuronal activities in 
      response to stimuli. It has been used for years to study odorant-induced 
      responses in insects (i.e., honeybees, Drosophila, and moths) and was recently 
      introduced into mosquitoes. Traditionally, calcium imaging in mosquitoes was 
      performed using nonspecific calcium indicator dyes to examine neuronal responses 
      in whole insect brain regions, but the development of genetically encoded calcium 
      indicators (GECIs) has facilitated the ability to perform functional calcium 
      imaging on specific tissues. For example, by specifically expressing a GECI in 
      olfactory neurons, the odor-induced responses of these neurons in peripheral 
      organs can be examined. Calcium imaging of mosquito antennae further provides an 
      advantageous method for simultaneously visualizing the activity of several 
      antennal neurons in a single experiment. In this protocol, we describe a calcium 
      imaging method to study odor-evoked responses in Anopheles coluzzii antennae 
      expressing the calcium indicator GCaMP6f. This method requires imaging equipment 
      (compound microscope, light sources, and camera), an odorant delivery system, and 
      image acquisition software. The mosquito preparation is straightforward but 
      requires practice to minimize mosquito movement during imaging. Recorded videos 
      can be analyzed using Fiji software to generate heatmaps and activity traces for 
      odorant-evoked responses. This protocol can also be used, with some 
      modifications, to study other peripheral organs (such as labella, palps, and 
      tarsi).
CI  - (c) 2022 Cold Spring Harbor Laboratory Press.
FAU - Afify, Ali
AU  - Afify A
AD  - The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland 21205, USA.
FAU - Potter, Christopher J
AU  - Potter CJ
AUID- ORCID: 0000-0002-5223-8112
AD  - The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School 
      of Medicine, Baltimore, Maryland 21205, USA cpotter@jhmi.edu.
LA  - eng
GR  - R01 AI137078/AI/NIAID NIH HHS/United States
PT  - Journal Article
DEP - 20221101
PL  - United States
TA  - Cold Spring Harb Protoc
JT  - Cold Spring Harbor protocols
JID - 101524530
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - *Anopheles/physiology
MH  - Calcium
MH  - Smell/physiology
MH  - Odorants
MH  - Drosophila
EDAT- 2022/08/13 06:00
MHDA- 2022/11/04 06:00
CRDT- 2022/08/12 12:32
PHST- 2022/08/13 06:00 [pubmed]
PHST- 2022/11/04 06:00 [medline]
PHST- 2022/08/12 12:32 [entrez]
AID - pdb.prot107918 [pii]
AID - 10.1101/pdb.prot107918 [doi]
PST - epublish
SO  - Cold Spring Harb Protoc. 2022 Nov 1;2022(11):Pdb.prot107918. doi: 
      10.1101/pdb.prot107918.