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Treatment procedure for ciliobrevin-D to the olfactory pits. 1. Fish were anesthetized <t>with</t> <t>ice-cold</t> water, and hold it upright position. 2. olfactory pits were indicated with arrows. 3. 40 µM ciliobrevin-D in 2 % agarose with 0.025% phenole red was air-injected into the posterior olfactory pit using a micropipette. 4.-5. After loading the ciliobrevin gel 6. Olfactory pits were covered by superglue 7. After curing (∼1 min), the fish were released in the fresh fish-conditioned water at ∼22°C, and their recovery was carefully observed until they actively swam. The superglue covers eventually fell off ∼30 min after curing.
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Treatment procedure for ciliobrevin-D to the olfactory pits. 1. Fish were anesthetized with ice-cold water, and hold it upright position. 2. olfactory pits were indicated with arrows. 3. 40 µM ciliobrevin-D in 2 % agarose with 0.025% phenole red was air-injected into the posterior olfactory pit using a micropipette. 4.-5. After loading the ciliobrevin gel 6. Olfactory pits were covered by superglue 7. After curing (∼1 min), the fish were released in the fresh fish-conditioned water at ∼22°C, and their recovery was carefully observed until they actively swam. The superglue covers eventually fell off ∼30 min after curing.

Journal: bioRxiv

Article Title: Novel mechanisms of chemosensory adaptation to the cave environment

doi: 10.64898/2026.04.03.716434

Figure Lengend Snippet: Treatment procedure for ciliobrevin-D to the olfactory pits. 1. Fish were anesthetized with ice-cold water, and hold it upright position. 2. olfactory pits were indicated with arrows. 3. 40 µM ciliobrevin-D in 2 % agarose with 0.025% phenole red was air-injected into the posterior olfactory pit using a micropipette. 4.-5. After loading the ciliobrevin gel 6. Olfactory pits were covered by superglue 7. After curing (∼1 min), the fish were released in the fresh fish-conditioned water at ∼22°C, and their recovery was carefully observed until they actively swam. The superglue covers eventually fell off ∼30 min after curing.

Article Snippet: The tissue around rosettes was carefully removed under the ice-cold RNase-free PBS (diethyl pyrocarbonate-treated PBS; ThermoFisher Scientific), washed once in the fresh ice-cold RNase-free PBS, recovered in the pre-weighed 1.5 mL tube (SP Bel-Art, Wayne, NJ, USA), and flash frozen by submerging it in liquid nitrogen.

Techniques: Olfactory, Injection