sNPF neuropeptide was administrated to IPCs and CC using a modified method from a previous study26 (link). sNPF was purchased from Thermo Fisher Scientific (sequence: AQRSPSLRLRF, purity > 95 %, unmodified). We made 50 mM stock solution of sNPF in DMSO (Sigma, D2650) and keep it at −80 °C. Pertussis toxin (Tocris, 3097), U73122 (Tocris, 1268), or U73343 (Tocris, 4133) was mixed with D-glucose or sNPF and treated to IPCs in the brain or AKH-producing cells in CC. We mixed 80 μM sNPF or 20 mM d- glucose contained AHL with 1 ng/μL Pertussis toxin (PT), 1 μM U73122, or 1 μM U73343 (a non-functional enantiomer of U73122). The concentrations of PT, U73122, and U73343 that were used in the experiments were determined based on the pilot experiments.
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Neuropeptide Regulation of Insulin-Producing Cells
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Variable analysis
independent variables
- SNPF neuropeptide administration
- Pertussis toxin (PT) treatment
- U73122 treatment
- U73343 treatment
- Response of IPCs and CC to sNPF, PT, U73122, and U73343 treatments
- SNPF stock solution preparation (50 mM in DMSO, stored at -80 °C)
- Concentrations of PT (1 ng/μL), U73122 (1 μM), and U73343 (1 μM) used in the experiments
- Positive control: sNPF neuropeptide treatment
- Negative control: U73343 (non-functional enantiomer of U73122) treatment
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