Tyrode solution contained (in mM): NaCl, 140; KCl, 5; CaCl2, 1; MgCl2, 1; NaHCO3, 5; HEPES, 10; Glucose, 10; sodium pyruvate, 10; pH adjusted to 7.4 with NaOH. Sucrose (500mM) was used as a sweet tastant. Recombinant murine leptin (20 ng/ml; PeproTech, Cat# 450-31), wortmannin (10 nM; Tocris Bioscience, Cat# 1,232), LY294002 (30 μM; Sigma-Aldrich, Cat# L9908), SHP099 (10 μM; Cayman Chemical, Cat# 20,000), and stattic (10 μM; Axon Medchem, Cat# 2,314) were applied from the basolateral side of TCs. Other chemicals were purchased from Wako Pure Chemical Industries or Nakarai Tesque. These concentrations of agents were selected as used in previous studies (Chen et al., 2016 (link); Mirshamsi et al., 2004 (link); Schust et al., 2006 (link); Yoshida et al., 2015 (link)).
Protocol detail
Tyrode Solution for Taste Cell Assay
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Corresponding Organization : Okayama University
Other organizations : Monell Chemical Senses Center, Tokyo Dental College, Kyushu University
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Variable analysis
independent variables
- Recombinant murine leptin (20 ng/ml; PeproTech, Cat# 450-31)
- Wortmannin (10 nM; Tocris Bioscience, Cat# 1,232)
- LY294002 (30 μM; Sigma-Aldrich, Cat# L9908)
- SHP099 (10 μM; Cayman Chemical, Cat# 20,000)
- Stattic (10 μM; Axon Medchem, Cat# 2,314)
- Sucrose (500mM) as a sweet tastant
- Not explicitly mentioned
- Tyrode solution: NaCl (140 mM), KCl (5 mM), CaCl2 (1 mM), MgCl2 (1 mM), NaHCO3 (5 mM), HEPES (10 mM), Glucose (10 mM), Sodium pyruvate (10 mM), pH adjusted to 7.4 with NaOH
- Positive control: Not specified
- Negative control: Not specified
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