PMID- 38148168
OWN - NLM
STAT- Publisher
LR  - 20240209
IS  - 1559-6095 (Electronic)
IS  - 1559-6095 (Linking)
DP  - 2024 Feb 9
TI  - Whole-Brain Calcium Imaging in Drosophila during Sleep and Wake.
LID - 10.1101/pdb.prot108419 [doi]
AB  - Genetically encoded calcium indicators (GECIs) allow for the noninvasive 
      evaluation of neuronal activity in vivo, and imaging GECIs in Drosophila has 
      become commonplace for understanding neural functions and connectivity in this 
      system. GECIs can also be used as read-outs for studying sleep in this model 
      organism. Here, we describe a methodology for tracking the activity of neurons in 
      the fly brain using a two-photon (2p) microscopy system. This method can be 
      adapted to perform functional studies of neural activity in Drosophila under both 
      spontaneous and evoked conditions, as well as during spontaneous or induced 
      sleep. We first describe a tethering and surgical procedure that allows survival 
      under the microscopy conditions required for long-term recordings. We then 
      outline the steps and reagents required for optogenetic activation of 
      sleep-promoting neurons while simultaneously recording neural activity from the 
      fly brain. We also describe the procedure for recording from two different 
      locations-namely, the top of the head (e.g., to record mushroom body calyx 
      activity) or the back of the head (e.g., to record central complex activity). We 
      also provide different strategies for recording from GECIs confined to the cell 
      body versus the entire neuron. Finally, we describe the steps required for 
      analyzing the multidimensional data that can be acquired. In all, this protocol 
      shows how to perform calcium imaging experiments in tethered flies, with a focus 
      on acquiring spontaneous and induced sleep data.
CI  - (c) 2023 Cold Spring Harbor Laboratory Press.
FAU - Tainton-Heap, Lucy
AU  - Tainton-Heap L
AD  - Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 
      4072, Australia.
FAU - Troup, Michael
AU  - Troup M
AD  - Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 
      4072, Australia.
FAU - Van De Poll, Matthew
AU  - Van De Poll M
AD  - Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 
      4072, Australia.
FAU - van Swinderen, Bruno
AU  - van Swinderen B
AD  - Queensland Brain Institute, The University of Queensland, Brisbane, Queensland 
      4072, Australia b.vanswinderen@uq.edu.au.
LA  - eng
PT  - Journal Article
DEP - 20240209
PL  - United States
TA  - Cold Spring Harb Protoc
JT  - Cold Spring Harbor protocols
JID - 101524530
SB  - IM
EDAT- 2023/12/27 00:41
MHDA- 2023/12/27 00:41
CRDT- 2023/12/26 21:23
PHST- 2023/12/27 00:41 [pubmed]
PHST- 2023/12/27 00:41 [medline]
PHST- 2023/12/26 21:23 [entrez]
AID - pdb.prot108419 [pii]
AID - 10.1101/pdb.prot108419 [doi]
PST - aheadofprint
SO  - Cold Spring Harb Protoc. 2024 Feb 9. doi: 10.1101/pdb.prot108419.