Current study, Van Den Herrewegen et al. (2021) | Adamsky et al. [36] | Nam et al. [16] | |
---|---|---|---|
Viral vectors and used chemogenetic tools | |||
Viral vector with DREADD construct |
*Gq-DREADD: AAV8-GFAP-hM3Dq-mCherry *Gi-DREADD: AAV8-GFAP-hM4Di-mCherry *Control vector: AAV8-GFAP-mCherry |
*Gq-DREADD: AAV8-GFAP-hM3Dq-mCherry *Gi-DREADD: / *Control vector: AAV8-GFAP-mCherry |
*Gq-DREADD: / *Gi-DREADD: AAV-GFAP-hM4Di-mCherry *Control vector: AAV-GFAP-GFP |
Animals | |||
Mouse strain/age | C57Bl6J/11–13 weeks | C57Bl6J/11–13 weeks | C57Bl6J/7–8 weeks |
Gender | Male | Male | Male |
Group/single housed | Group | Group | Ns |
Slicing conditions | |||
Slice orientation and thickness |
Coronal 400 µm |
Coronal 400 µm |
Transverse 350–400 µm |
Slicing solution (in mM) | 75 Sucrose, 87 NaCl, 2.5 KCl, 25 NaHCO3, 1.25 NaH2PO4, 7 MgCl2, 0.5 CaCl2, 0.6 Ascorbic acid and 25 glucose | 126 NaCl, 2.6 KCl, 26 NaHCO3, 1.25 NaH2PO4, 1 MgCl2, 0.625 CaCl2 and 10 glucose | 212.7 sucrose, 5 KCl, 26 NaHCO3, 1.23 NaH2PO4, 10 MgSO4, 0.5 CaCl2, and 10 dextrose |
Slice recovery conditions | 1 h submerged | 1 h submerged | 1 h ns |
DREADD agonist | |||
DREADD agonist | CNO | CNO | CNO |
Concentration | 10 µM | 10 µM | ns |
Duration of CNO application | 25 min | 25 min | 10 min |
Slice electrophysiology recording conditions | |||
Stimulation frequency | 0.033 Hz | 0.017 Hz | 0.07 Hz |
Stimulations during baseline | 30 min (60 stimulations) | 10 min (10 stimulations) | 10 min (40 stimulations) |
Pause in stimulation | Yes | Yes | No |
Stimulations necessary for induction of synaptic potentiation |
Gq-DREADD: 21 Gi-DREADD: 1 |
Gq-DREADD: 1 (1 mM Mg2+) 21 (2 mM Mg2+) | Gi-DREADD: 40+ additional 40 Hz stimulus |
Bath temperature | 28 °C | 32 °C | 28–30 °C |
aCSF composition (in mM) | 124 NaCl, 3 KCl, 26 NaHCO3, 1.4 NaH2PO4, 1 MgSO4, 2 CaCl2 and 10 glucose | 126 NaCl, 2.6 KCl, 26 NaHCO3, 1.25 NaH2PO4, 1 MgCl2, 2 CaCl2 and 10 glucose | 124 NaCl, 5 KCl, 26 NaHCO3, 1.23 NaH2PO4, 1 MgSO4, 2 CaCl2 and 10 dextrose |