PMID- 34633379
OWN - NLM
STAT- MEDLINE
DCOM- 20211021
LR  - 20211021
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 175
DP  - 2021 Sep 21
TI  - Simultaneous Calcium Imaging and Glucose Stimulation in Living Zebrafish to 
      Investigate In Vivo beta-Cell Function.
LID - 10.3791/62347 [doi]
AB  - The pancreatic beta-cells sustain systemic glucose homeostasis by producing and 
      secreting insulin according to the blood glucose levels. Defects in beta-cell 
      function are associated with hyperglycemia that can lead to diabetes. During the 
      process of insulin secretion, beta-cells experience an influx of Ca(2+). Thus, 
      imaging the glucose-stimulated Ca(2+) influx using genetically encoded calcium 
      indicators (GECIs) provides an avenue to studying beta-cell function. Previously, 
      studies showed that isolated zebrafish islets expressing GCaMP6s exhibit 
      significant Ca(2+) activity upon stimulation with defined glucose concentrations. 
      However, it is paramount to study how beta-cells respond to glucose not in 
      isolation, but in their native environment, where they are systemically 
      connected, vascularized, and densely innervated. To this end, the study leveraged 
      the optical transparency of the zebrafish larvae at early stages of development 
      to illuminate beta-cell activity in vivo. Here, a detailed protocol for Ca(2+) 
      imaging and glucose stimulation to investigate beta-cell function in vivo is 
      presented. This technique allows to monitor the coordinated Ca(2+) dynamics in 
      beta-cells with single-cell resolution. Additionally, this method can be applied to 
      work with any injectable solution such as small molecules or peptides. 
      Altogether, the protocol illustrates the potential of the zebrafish model to 
      investigate islet coordination in vivo and to characterize how environmental and 
      genetic components might affect beta-cell function.
FAU - Delgadillo-Silva, Luis Fernando
AU  - Delgadillo-Silva LF
AD  - Centre for Regenerative Therapies TU Dresden.
FAU - Akhtar, Mohammad Nadeem
AU  - Akhtar MN
AD  - Centre for Regenerative Therapies TU Dresden.
FAU - Tasoz, Emirhan
AU  - Tasoz E
AD  - Centre for Regenerative Therapies TU Dresden.
FAU - Ninov, Nikolay
AU  - Ninov N
AD  - Centre for Regenerative Therapies TU Dresden; German Center for Diabetes Research 
      (DZD e.V.), Paul Langerhans Institute Dresden of the Helmholtz Center Munich at 
      the University Hospital Carl Gustav Carus of TU Dresden; 
      nikolay.ninov@tu-dresden.de.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20210921
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - IY9XDZ35W2 (Glucose)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - Animals
MH  - *Calcium/metabolism
MH  - Glucose/metabolism
MH  - Insulin Secretion
MH  - *Insulin-Secreting Cells/metabolism
MH  - Zebrafish/metabolism
EDAT- 2021/10/12 06:00
MHDA- 2023/02/25 06:00
CRDT- 2021/10/11 12:21
PHST- 2021/10/11 12:21 [entrez]
PHST- 2021/10/12 06:00 [pubmed]
PHST- 2023/02/25 06:00 [medline]
AID - 10.3791/62347 [doi]
PST - epublish
SO  - J Vis Exp. 2021 Sep 21;(175). doi: 10.3791/62347.