Table 1.
Summary of animals used for the experiments.
| Procedure | Virus/plasmid type | Animals | Injection location | Sample size | Time after procedure (sample size) | Time unit |
|---|---|---|---|---|---|---|
| Electroporation, in vivo | pPB.CAG.GFP | Tadpoles | OB/T/OT | 11 | 16 | h |
| pCS2.EGFP(CAAX) | Tadpoles | OB/T/OT | 7 | 16 | h | |
| pCS2FA.ChR2.YFP | Tadpoles | OB/T/OT | 13 | 24 | h | |
| pCS2FA.ChR2.YFP | Frogs | OB/T/OT | 8 | 5 (2), 6 (6) | h | |
| Electroporation, in vitro | pPB.CAG.GFP | Frogs | OB/T/OT | 1 | 27 | h |
| pCS2.EGFP(CAAX) | Frogs | OB/T/OT | 5 | 16 (3), 22 (2) | h | |
| pCS2FA.ChR2.YFP | Frogs | OB/T/OT | 5 | 2 (1), 3 (3), 4 (1) | d | |
| AAV injection, in vivo | AAV9.hSyn.hChR2(H134R)eYFP.WPRE.hGH | Frogs | OB/T/OT | 34 | 21 | d |
| retroAAV mCherry-Cre | Frogs | OB/T/OT | 8 | 21 | d | |
| AAV2/1.Syn.ChR2(H134R)eYFP.AWP.hGH | Frogs | OB/T/OT | 3 | 21 | d | |
| AAV-PHPeB:Cag-mNeonGreen | Frogs | OB/T/OT | 4 | 21 | d | |
| LV injection, in vivo | LV-SIN-CMV-eGFP | Frogs | OB/T/OT | 4 | 6 | w |
| EIAV-TLoop-ChR2-YFP | Frogs | OB/T/OT | 9 | 12 | w | |
| EIAV-TLoop-ChR2-YFP | Tadpoles | OB/T/OT | 9 | 12 | d | |
| VSV injection, in vivo | VSV(VSV-G) plaque 14 | Frogs | OB/T/OT | 8 | 2 (4), 4 (2), 6 (2) | d |
| VSV(VSV-G) plaque 14 | Frogs | n.IX-X | 6 | 2 (2), 5 (1), 6 (1), 7 (2) | d | |
| VSV(VSV-G) plaque 21 | Frogs | OB/T/OT | 28 | 2 (10), 4 (3), 7 (4), 10 (11) | d | |
| VSV(VSV-G) plaque 21 | Frogs | n.IX-X | 18 | 3 | d | |
| VSV(VSV-G) plaque 21 | Frogs | nerve IX-X | 7 | 2 (1), 28 (4), 35 (2) | d | |
| VSV(VSV-G) plaque 21 | Tadpoles | OB/T/OT | 42 | 1 (9), 2 (26), 4 (4), 5 (3) | d | |
| VSV(VSV-G) plaque 21 | Tadpoles | nose | 85 | 1 (2), 2 (29), 3 (1), 4 (22), 5 (31) | d | |
| VSV(RABV-G) | Frogs | OB/T/OT | 7 | 1 (1), 2 (4), 3 (2) | d | |
| VSV(RABV-G) | Frogs | n.IX-X | 8 | 2 (3), 8 (1), 9 (2), 10 (2) | d | |
| VSV(RABV-G) | Frogs | laryngeal muscles | 10 | 21 (1), 22 (1), 23 (4), 27 (2), 28 (2) | d | |
| VSV(RABV-G) | Frogs | gastrocnemius muscles | 4 | 7 | d | |
| VSV(RABV-G) | Tadpoles | OB/T/OT | 12 | 3 | d | |
| G-Deleted VSV-tdTomato | Frogs | OB/T/OT | 3 | 2 | d | |
| G-Deleted VSV-tdTomato | Frogs | n.IX-X | 1 | 2 | d | |
| G-Deleted VSV-ChR2YFP | Frogs | OB/T/OT | 7 | 2 (4), 5 (3) | d | |
| G-Deleted VSV-ChR2YFP | Frogs | n.IX-X | 2 | 9 (1), 10 (1) | d |
Injection site indicates the location where plasmid or virus was injected. “Time after procedure” column indicates the interval between the plasmid electroporation/viral injection and the time when brain was fixed, indicated in hours (h), days (d), or weeks (w) in the “time unit” column. When animals in an experimental group had variable amount of survival time, the number of animals sacrificed at each time point is shown in parenthesis in “Time after procedure” column. Injection location within the brain was separated into region rich in cell bodies including olfactory bulb, telencephalon, and optic tectum (OB/T/OT) and the region rich in myelinated axons, the laryngeal motor nucleus in the brainstem (n.IX-X). AAV, adeno-associated virus; LV, lentivirus, OB/T/OT, olfactory bulb/telencephalon/optic tectum; n.IX-X, laryngeal motor nucleus; VSV, vesicular stomatitis virus.